Descriptor: "MESH: Structure-Activity Relationship" - Searchworks@Jio Institute Digital Library Search Results

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364 results on '"MESH: Structure-Activity Relationship"'

1. Shedding X-ray Light on the Role of Magnesium in the Activity of Mycobacterium tuberculosis Salicylate Synthase (MbtI) for Drug Design

Author: Arianna Gelain, Fiorella Meneghetti, Josè Camilla Sammartino, Giulio Poli, Elena Pini, Stefania Villa, Laurent R. Chiarelli, Marco Bellinzoni, Giovanni Stelitano, Alessio Porta, Giangiacomo Beretta, Tiziano Tuccinardi, M. Mori, Università degli Studi di Milano [Milano] (UNIMI), Università degli Studi di Pavia, University of Pisa - Università di Pisa, Temple University [Philadelphia], Pennsylvania Commonwealth System of Higher Education (PCSHE), Microbiologie structurale - Structural Microbiology (Microb. Struc. (UMR_3528 / U-Pasteur_5)), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), This work was funded by University of Milan (Linea B) and the Italian Ministry of Education, University and Research (MIUR): Dipartimenti di Eccellenza Program (2018–2022) - Dept. of Biology and Biotechnology 'L. Spallanzani', University of Pavia. Partial support was also provided by institutional grants from Institut Pasteur and CNRS., Università degli Studi di Milano = University of Milan (UNIMI), Università degli Studi di Pavia = University of Pavia (UNIPV), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)
Subjects: Models, Molecular, Siderophore, MESH: Mycobacterium tuberculosis, Protein Conformation, MESH: Drug Design, Crystallography, X-Ray, 01 natural sciences, MESH: Protein Conformation, MESH: Structure-Activity Relationship, Models, MESH: Magnesium, Drug Discovery, Magnesium, Ternary complex, chemistry.chemical_classification, 0303 health sciences, Crystallography, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], biology, ATP synthase, 3. Good health, Biochemistry, Molecular Medicine, MESH: Models, Molecular, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Lyases, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Cofactor, Article, Mycobacterium tuberculosis, 03 medical and health sciences, Structure-Activity Relationship, [CHIM.CRIS]Chemical Sciences/Cristallography, Structure–activity relationship, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, 030304 developmental biology, Rational design, Molecular, MESH: Crystallography, X-Ray, biology.organism_classification, MESH: Lyases, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, Enzyme, chemistry, Drug Design, X-Ray, biology.protein, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM]
Abstract: International audience; The Mg2+-dependent Mycobacterium tuberculosis salicylate synthase (MbtI) is a key enzyme involved in the biosynthesis of siderophores. Because iron is essential for the survival and pathogenicity of the microorganism, this protein constitutes an attractive target for antitubercular therapy, also considering the absence of homologous enzymes in mammals. An extension of the structure-activity relationships of our furan-based candidates allowed us to disclose the most potent competitive inhibitor known to date (10, Ki = 4 μM), which also proved effective on mycobacterial cultures. By structural studies, we characterized its unexpected Mg2+-independent binding mode. We also investigated the role of the Mg2+ cofactor in catalysis, analyzing the first crystal structure of the MbtI-Mg2+-salicylate ternary complex. Overall, these results pave the way for the development of novel antituberculars through the rational design of improved MbtI inhibitors.
Published: 2020

2. Fragment-Based Phenotypic Lead Discovery To Identify New Drug Seeds That Target Infectious Diseases

Author: Paul Ogadinma, Yann Ayotte, Laurent Chatel-Chaix, François Bilodeau, Marthe Lebughe, Mena Cimino, Giulia Manina, Aïssatou Aïcha Sow, Frédéric J. Veyrier, Albert Descoteaux, Eve Bernet, Dominic Gagnon, Sarah-Lisa Ouali, Maxime Mistretta, Dave Richard, Steven R. LaPlante, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), NMX Research and Solutions Inc., Individualité microbienne et infection - Microbial Individuality and Infection, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Centre de recherche du CHU de Québec-Université Laval (CRCHUQ), CHU de Québec–Université Laval, and Université Laval [Québec] (ULaval)-Université Laval [Québec] (ULaval)
Subjects: biology, [SDV]Life Sciences [q-bio], MESH: Anti-Infective Agents, Drug Evaluation, Preclinical, Plasmodium falciparum, General Medicine, Computational biology, biology.organism_classification, Leishmania, Biochemistry, Small molecule, Phenotype, Structure-Activity Relationship, MESH: Structure-Activity Relationship, MESH: Drug Discovery, Anti-Infective Agents, Drug Discovery, Nucleic acid, Molecular Medicine, Bioassay, MESH: Drug Evaluation, Preclinical, Neisseria, Mycobacterium
Abstract: International audience; Fragment-based lead discovery has emerged over the last decades as one of the most powerful techniques for identifying starting chemical matter to target specific proteins or nucleic acids in vitro. However, the use of such low-molecular-weight fragment molecules in cell-based phenotypic assays has been historically avoided because of concerns that bioassays would be insufficiently sensitive to detect the limited potency expected for such small molecules and that the high concentrations required would likely implicate undesirable artifacts. Herein, we applied phenotype cell-based screens using a curated fragment library to identify inhibitors against a range of pathogens including Leishmania, Plasmodium falciparum, Neisseria, Mycobacterium, and flaviviruses. This proof-of-concept shows that fragment-based phenotypic lead discovery (FPLD) can serve as a promising complementary approach for tackling infectious diseases and other drug-discovery programs.
Published: 2021

3. Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation

Author: Solène N Lefebvre, Pierre-Jean Corringer, Anaïs Menny, Antoine Taly, Karima Medjebeur, Récepteurs Canaux - Channel Receptors, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Collège doctoral [Sorbonne universités], Sorbonne Université (SU), Laboratoire de biochimie théorique [Paris] (LBT (UPR_9080)), Institut de biologie physico-chimique (IBPC (FR_550)), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Centre National de la Recherche Scientifique (CNRS), ANR-13-BSV8-0020,Pentagate,Mécanismes d'activation et de désensibilisation d'un récepteur-canal pentamérique(2013), ANR-11-LABX-0011,DYNAMO,Dynamique des membranes transductrices d'énergie : biogénèse et organisation supramoléculaire.(2011), European Project: 788974,H2020 | ERC | ERC-ADG,Dynacotine(2019), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Collège Doctoral, Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), The work was supported by the ‘Agence Nationale de la Recherche’ (Grant ANR-13-BSV8-0020, Pentagate), the doctoral school ED3C and the ‘Foundation pour la Recherche Médicale’ (PhD funding to SNL), the ‘Initiative d'Excellence’ (cluster of excellence LABEX Dynamo, ANR-11-LABX-0011 to AT) and the ERC (Grant no. 788974, Dynacotine)., and Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)
Subjects: Time Factors, Protein Conformation, [SDV]Life Sciences [q-bio], Structural Biology and Molecular Biophysics, Xenopus, DNA Mutational Analysis, Gating, MESH: Structure-Activity Relationship, MESH: Protein Conformation, 0302 clinical medicine, Protein structure, structural biology, intermediate conformation, MESH: DNA Mutational Analysis, Biology (General), MESH: Bacterial Proteins, 0303 health sciences, MESH: Escherichia coli, Chemistry, General Neuroscience, MESH: Cyanobacteria, General Medicine, Molecular Docking Simulation, Transmembrane domain, MESH: Ligand-Gated Ion Channels, Medicine, Ion Channel Gating, Research Article, MESH: Mutation, pentameric ligand-gated ion channels, QH301-705.5, Protein subunit, Science, GLIC, Allosteric regulation, fluorescence quenching, Cyanobacteria, Models, Biological, General Biochemistry, Genetics and Molecular Biology, Structure-Activity Relationship, 03 medical and health sciences, Bacterial Proteins, molecular biophysics, MESH: Molecular Docking Simulation, Escherichia coli, Ion channel, 030304 developmental biology, General Immunology and Microbiology, MESH: Time Factors, MESH: Models, Biological, E. coli, MESH: Ion Channel Gating, Ligand-Gated Ion Channels, allosteric coupling, Structural biology, Mutation, Biophysics, 030217 neurology & neurosurgery, Neuroscience
Abstract: International audience; Pentameric ligand-gated ion channels (pLGICs) mediate chemical signaling through a succession of allosteric transitions that are yet not completely understood as intermediate states remain poorly characterized by structural approaches. In a previous study on the prototypic bacterial proton-gated channel GLIC, we generated several fluorescent sensors of the protein conformation that report a fast transition to a pre-active state, which precedes the slower process of activation with pore opening. Here, we explored the phenotype of a series of allosteric mutations, using paralleled steady-state fluorescence and electrophysiological measurements over a broad pH range. Our data, fitted to a 3-states Monod-Wyman-Changeux (MWC) model, show that mutations at the subunit interface in the extracellular domain (ECD) principally alter pre-activation, while mutations in the lower ECD and in the transmembrane domain principally alter activation. We also show that propofol alters both transitions. Data are discussed in the framework of transition pathways generated by normal mode analysis (iModFit) that suggest collective protein motions concerted with pore opening. It further supports that pre-activation involves major quaternary compaction of the ECD, and suggests that activation involves principally a re-organization of a 'central gating region' involving a contraction of the ECD β-sandwich and the tilt of the channel lining M2 helix.
Published: 2021

4. Focus on DNA Glycosylases—A Set of Tightly Regulated Enzymes with a High Potential as Anticancer Drug Targets

Author: Chandini Bhaskar Naidu, Joanna Timmins, Fabienne Hans, Muge Senarisoy, Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)
Subjects: DNA Repair, Review, lcsh:Chemistry, chemistry.chemical_compound, 0302 clinical medicine, MESH: Structure-Activity Relationship, Neoplasms, MESH: Molecular Targeted Therapy, inhibitors, MESH: Animals, MESH: Neoplasms, Molecular Targeted Therapy, MESH: DNA Glycosylases, lcsh:QH301-705.5, Spectroscopy, chemistry.chemical_classification, MESH: DNA Repair, 0303 health sciences, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MESH: Gene Expression Regulation, Enzymologic, General Medicine, Base excision repair, 3. Good health, Computer Science Applications, DNA glycosylases, 030220 oncology & carcinogenesis, Disease Susceptibility, MESH: Enzyme Activation, DNA repair, MESH: Disease Susceptibility, Antineoplastic Agents, Biology, base excision repair, Catalysis, Gene Expression Regulation, Enzymologic, Inorganic Chemistry, 03 medical and health sciences, Structure-Activity Relationship, medicine, Animals, Humans, cancer, Physical and Theoretical Chemistry, Molecular Biology, 030304 developmental biology, MESH: DNA Damage, MESH: Humans, drug resistance, Mechanism (biology), Organic Chemistry, Cancer, medicine.disease, Enzyme Activation, Enzyme, chemistry, lcsh:Biology (General), lcsh:QD1-999, DNA glycosylase, Cancer cell, Cancer research, MESH: Antineoplastic Agents, DNA, DNA Damage
Abstract: International audience; Cancer is the second leading cause of death with tens of millions of people diagnosed with cancer every year around the world. Most radio- and chemotherapies aim to eliminate cancer cells, notably by causing severe damage to the DNA. However, efficient repair of such damage represents a common mechanism of resistance to initially effective cytotoxic agents. Thus, development of new generation anticancer drugs that target DNA repair pathways, and more particularly the base excision repair (BER) pathway that is responsible for removal of damaged bases, is of growing interest. The BER pathway is initiated by a set of enzymes known as DNA glycosylases. Unlike several downstream BER enzymes, DNA glycosylases have so far received little attention and the development of specific inhibitors of these enzymes has been lagging. Yet, dysregulation of DNA glycosylases is also known to play a central role in numerous cancers and at different stages of the disease, and thus inhibiting DNA glycosylases is now considered a valid strategy to eliminate cancer cells. This review provides a detailed overview of the activities of DNA glycosylases in normal and cancer cells, their modes of regulation, and their potential as anticancer drug targets.
Published: 2020

5. Design, synthesis, and biological evaluation of dual targeting inhibitors of histone deacetylase 6/8 and bromodomain BRPF1

Author: Manfred Jung, Karin Schmidtkunz, Elizabeth R Morales, Stefan Günther, Wolfgang Sippl, Martin Hügle, Frank Erdmann, Patrik Zeyen, Ehab Ghazy, Dina Robaa, Christophe Romier, Matthias Schmidt, Daniel Herp, Martin-Luther-Universität Halle Wittenberg (MLU), Albert-Ludwigs-Universität Freiburg, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Romier, Christophe
Subjects: Bromodomain, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, MESH: Drug Design, Histone Deacetylase 6, 01 natural sciences, MESH: Structure-Activity Relationship, Drug Discovery, MESH: Molecular Dynamics Simulation, MESH: Histone Deacetylase Inhibitors, 0303 health sciences, biology, Chemistry, Acetylation, General Medicine, Hydroxamic acids, 3. Good health, Cell biology, DNA-Binding Proteins, Molecular Docking Simulation, Leukemia, Myeloid, Acute, Histone, MESH: Repressor Proteins, Epigenetics, MESH: Leukemia, Myeloid, Acute, MESH: Acetylation, MESH: Cell Line, Tumor, [SDV.SP.MED] Life Sciences [q-bio]/Pharmaceutical sciences/Medication, [CHIM.THER] Chemical Sciences/Medicinal Chemistry, Antineoplastic Agents, [SDV.CAN]Life Sciences [q-bio]/Cancer, Molecular Dynamics Simulation, Histone Deacetylases, 03 medical and health sciences, Structure-Activity Relationship, [SDV.SP.MED]Life Sciences [q-bio]/Pharmaceutical sciences/Medication, [SDV.CAN] Life Sciences [q-bio]/Cancer, Cell Line, Tumor, MESH: Molecular Docking Simulation, Humans, Dual targeting inhibitors, 030304 developmental biology, Adaptor Proteins, Signal Transducing, MESH: Adaptor Proteins, Signal Transducing, Pharmacology, Acute myeloid leukemia, MESH: Humans, 010405 organic chemistry, Organic Chemistry, HDAC8, HDAC6, 0104 chemical sciences, Histone Deacetylase Inhibitors, Repressor Proteins, MESH: Histone Deacetylases, Docking (molecular), PHD finger, BRPF1, Drug Design, biology.protein, MESH: Antineoplastic Agents, MESH: Histone Deacetylase 6, MESH: DNA-Binding Proteins
Abstract: International audience; Histone modifying proteins, specifically histone deacetylases (HDACs) and bromodomains, have emerged as novel promising targets for anticancer therapy. In the current work, based on available crystal structures and docking studies, we designed dual inhibitors of both HDAC6/8 and the bromodomain and PHD finger containing protein 1 (BRPF1). Biochemical and biophysical tests showed that compounds 23a,b and 37 are nanomolar inhibitors of both target proteins. Detailed structure-activity relationships were deduced for the synthesized inhibitors which were supported by extensive docking and molecular dynamics studies. Cellular testing in acute myeloid leukemia (AML) cells showed only a weak effect, most probably because of the poor permeability of the inhibitors. We also aimed to analyse the target engagement and the cellular activity of the novel inhibitors by determining the protein acetylation levels in cells by western blotting (tubulin vs histone acetylation), and by assessing their effects on various cancer cell lines.
Published: 2020

6. Breakthroughs in medicinal chemistry: new targets and mechanisms, new drugs, new hopes-7

Author: Gütschow, Michael, Eynde, Jean Jacques Vanden, Jampilek, Josef, Kang, CongBao, Mangoni, Arduino, Fossa, Paola, Karaman, Rafik, Trabocchi, Andrea, Scott, Peter, Reynisson, Jóhannes, Rapposelli, Simona, Galdiero, Stefania, Winum, Jean-Yves, Brullo, Chiara, Prokai-Tatrai, Katalin, Sharma, Arun, Schapira, Matthieu, Azuma, Yasu-Taka, Cerchia, Laura, Spetea, Mariana, Torri, Giangiacomo, Collina, Simona, Geronikaki, Athina, García-Sosa, Alfonso, Vasconcelos, M Helena, Sousa, Maria Emília, Kosalec, Ivan, Tuccinardi, Tiziano, Duarte, Iola, Salvador, Jorge, Bertinaria, Massimo, Pellecchia, Maurizio, Amato, Jussara, Rastelli, Giulio, Gomes, Paula, Guedes, Rita, Sabatier, Jean-Marc, Estévez-Braun, Ana, Pagano, Bruno, Mangani, Stefano, Ragno, Rino, Kokotos, George, Brindisi, Margherita, González, Florenci, Borges, Fernanda, Miloso, Mariarosaria, Rautio, Jarkko, Muñoz-Torrero, Diego, Vanden Eynde, Jean Jacques, Vasconcelos, M. Helena, Gutschow, M., Eynde, J. J. V., Jampilek, J., Kang, C., Mangoni, A. A., Fossa, P., Karaman, R., Trabocchi, A., Scott, P. J. H., Reynisson, J., Rapposelli, S., Galdiero, S., Winum, J. -Y., Brullo, C., Prokai-Tatrai, K., Sharma, A. K., Schapira, M., Azuma, Y. -T., Cerchia, L., Spete, M., Torri, G., Collina, S., Geronikaki, A., Garcia-Sosa, A. T., Helena Vasconcelos, M., Sousa, M. E., Kosalec, I., Tuccinardi, T., Duarte, I. F., Salvador, J. A. R., Bertinaria, M., Pellecchia, M., Amato, J., Rastelli, G., Gomes, P. A. C., Guedes, R. C., Sabatier, J. -M., Estevez-Braun, A., Pagano, B., Mangani, S., Ragno, R., Kokotos, G., Brindisi, M., Gonzalez, F. V., Borges, F., Miloso, M., Rautio, J., Munoz-Torrero, D., Institut de neurophysiopathologie (INP), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Gutschow, M, Eynde, J, Jampilek, J, Kang, C, Mangoni, A, Fossa, P, Karaman, R, Trabocchi, A, Scott, P, Reynisson, J, Rapposelli, S, Galdier, S, Winum, J, Brullo, C, Prokai-Tatrai, K, Sharma, A, Schapira, M, Azuma, Y, Cerchia, L, Spete, M, Torri, G, Collina, S, Geronikaki, A, Garcia-Sosa, A, Helena Vasconcelos, M, Sousa, M, Kosalec, I, Tuccinardi, T, Duarte, I, Salvador, J, Bertinaria, M, Pellecchia, M, Amato, J, Rastelli, G, Gomes, P, Guedes, R, Sabatier, J, Estevez-Braun, A, Pagano, B, Mangani, S, Ragno, R, Kokotos, G, Brindisi, M, Gonzalez, F, Borges, F, Miloso, M, Rautio, J, and Munoz-Torrero, D
Subjects: RM, Pharmaceutical research, molecular targeted therapy, Chemistry, Pharmaceutical, MESH: Pharmaceutical Preparations, [SDV]Life Sciences [q-bio], Pharmaceutical Science, animals, chemistry, pharmaceutical, drug discovery, humans, pharmaceutical preparations, structure-activity relationship, Q1, chemistry, 01 natural sciences, Clinical chemistry, Analytical Chemistry, lcsh:QD241-441, 03 medical and health sciences, MESH: Chemistry, Pharmaceutical, Química clínica, MESH: Structure-Activity Relationship, lcsh:Organic chemistry, MESH: Drug Discovery, CHIM/06 - CHIMICA ORGANICA, MESH: Molecular Targeted Therapy, pharmaceutical, MESH: Animals, RM695, Investigació farmacèutica, Physical and Theoretical Chemistry, 030304 developmental biology, 0303 health sciences, MESH: Humans, 010405 organic chemistry, Medicinal Chemistry, New Targets, New Mechanisms, New Drugs, Organic Chemistry, R735, R1, 3. Good health, 0104 chemical sciences, Editorial, n/a, Chemistry (miscellaneous), Molecular Medicine, Breakthroughs, Medicinal Chemistry, medicinal chemistry, targets, drugs, molecules, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: Breakthroughs in Medicinal Chemistry: New Targets and Mechanisms, New Drugs, New Hopes is a series of editorials which is published on a biannual basis by the Editorial Board of the Medicinal Chemistry section of the journal Molecules. In these editorials, we highlight in brief reports (of about one hundred words) a number of recently published articles that describe crucial findings, such as the discovery of novel drug targets and mechanisms of action or novel classes of drugs, which may inspire future medicinal chemistry endeavors devoted to addressing prime unmet medical needs.
Published: 2020

7. Structures of MERS-CoV spike glycoprotein in complex with sialoside attachment receptors

Author: Wentao Li, Alexandra C. Walls, M. Alejandra Tortorici, Z. Wang, Maximilian M. Sauer, Young-Jun Park, Frank DiMaio, David Veesler, Berend Jan Bosch, Department of Biochemistry [Washington ], University of Washington [Seattle], Utrecht University [Utrecht], Virologie Structurale - Structural Virology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Research reported in this publication was supported by the National Institute of General Medical Sciences (R01GM120553, D.V.), the National Institute of Allergy and Infectious Diseases (HHSN272201700059C, D.V.), a Pew Biomedical Scholars Award (D.V.), an Investigators in the Pathogenesis of Infectious Disease Award from the Burroughs Wellcome Fund (D.V.) and a Swiss National Science Foundation PostDoc Mobility fellowship (M.M.S.). M.A.T. acknowledges support from the Institut Pasteur. This work was also supported by the Arnold and Mabel Beckman cryo-EM center at the University of Washington, dI&I I&I-1, LS Virologie, and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Viral membrane fusion, Models, Molecular, Protein Conformation, viruses, [SDV]Life Sciences [q-bio], MESH: Spike Glycoprotein, Coronavirus, Plasma protein binding, medicine.disease_cause, chemistry.chemical_compound, 0302 clinical medicine, Protein structure, MESH: Structure-Activity Relationship, MESH: Protein Conformation, Structural Biology, Models, Neuraminic acid, Protein Interaction Mapping, Carbohydrate Conformation, Viral, MESH: Carbohydrate Conformation, Receptor, chemistry.chemical_classification, 0303 health sciences, Middle East Respiratory Syndrome Coronavirus/chemistry, Spike Glycoprotein, 3. Good health, Cell biology, Coronavirus/chemistry, Spike Glycoprotein, Coronavirus, Middle East Respiratory Syndrome Coronavirus, MESH: Protein Domains, MESH: Cryoelectron Microscopy, MESH: Models, Molecular, Protein Binding, Viral protein, Dipeptidyl Peptidase 4, Coronacrisis-Taverne, MESH: Sialic Acids, Article, 03 medical and health sciences, Structure-Activity Relationship, Sialic Acids/chemistry, Protein Domains, Viral entry, MESH: Hemagglutination, Viral, medicine, Humans, MESH: Protein Binding, Binding site, Molecular Biology, Hemagglutination, Viral, 030304 developmental biology, Spike Glycoprotein, Coronavirus/chemistry, Binding Sites, MESH: Humans, Hemagglutination, Dipeptidyl Peptidase 4/chemistry, Cryoelectron Microscopy, MESH: Protein Interaction Mapping, MESH: Middle East Respiratory Syndrome Coronavirus, Molecular, MESH: Dipeptidyl Peptidase 4, chemistry, MESH: Binding Sites, Sialic Acids, Glycoprotein, 030217 neurology & neurosurgery
Abstract: The Middle East respiratory syndrome coronavirus (MERS-CoV) causes severe and often lethal respiratory illness in humans, and no vaccines or specific treatments are available. Infections are initiated via binding of the MERS-CoV spike (S) glycoprotein to sialosides and dipeptidyl-peptidase 4 (the attachment and entry receptors, respectively). To understand MERS-CoV engagement of sialylated receptors, we determined the cryo-EM structures of S in complex with 5-N-acetyl neuraminic acid, 5-N-glycolyl neuraminic acid, sialyl-LewisX, α2,3-sialyl-N-acetyl-lactosamine and α2,6-sialyl-N-acetyl-lactosamine at 2.7–3.0 Å resolution. We show that recognition occurs via a conserved groove that is essential for MERS-CoV S-mediated attachment to sialosides and entry into human airway epithelial cells. Our data illuminate MERS-CoV S sialoside specificity and suggest that selectivity for α2,3-linked over α2,6-linked receptors results from enhanced interactions with the former class of oligosaccharides. This study provides a structural framework explaining MERS-CoV attachment to sialoside receptors and identifies a site of potential vulnerability to inhibitors of viral entry., Cryo-EM structures of MERS-CoV S glycoprotein trimer in complex with different sialosides reveal how the virus engages with sialylated receptors, providing insight into receptor specificity and selectivity.
Published: 2019

8. Cell Biology of T Cell Receptor Expression and Regulation

Author: Balbino Alarcón, Andrés Alcover, Vincenzo Di Bartolo, Biologie Cellulaire des Lymphocytes - Lymphocyte Cell Biology, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], Centro de Biología Molecular Severo Ochoa [Madrid] (CBMSO), Universidad Autonoma de Madrid (UAM)-Consejo Superior de Investigaciones Científicas [Madrid] (CSIC), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Universidad Autónoma de Madrid (UAM)-Consejo Superior de Investigaciones Científicas [Madrid] (CSIC)
Subjects: MESH: Signal Transduction, 0301 basic medicine, MESH: CD3 Complex, CD3 Complex, T-Lymphocytes, recycling, MESH: Structure-Activity Relationship, Immunology and Allergy, MESH: Animals, biology, MESH: Receptors, Antigen, T-Cell, MESH: Gene Expression Regulation, Endocytosis, 3. Good health, Cell biology, medicine.anatomical_structure, [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, TCR-CD3, MESH: Endocytosis, Signal transduction, signaling, Signal Transduction, assembly, Endosome, T cell, Immunology, MESH: Immunomodulation, Receptors, Antigen, T-Cell, MESH: Proteolysis, Endosomes, Major histocompatibility complex, Immunomodulation, Structure-Activity Relationship, 03 medical and health sciences, conformational change, Antigen, medicine, Animals, Humans, MESH: Humans, Cell Membrane, T-cell receptor, MESH: Multiprotein Complexes, MESH: T-Lymphocytes, 030104 developmental biology, Gene Expression Regulation, MESH: Endosomes, Multiprotein Complexes, Proteolysis, MESH: Biomarkers, biology.protein, Biomarkers, MESH: Cell Membrane
Abstract: International audience; T cell receptors (TCRs) are protein complexes formed by six different polypeptides. In most T cells, TCRs are composed of αβ subunits displaying immunoglobulin-like variable domains that recognize peptide antigens associated with major histocompatibility complex molecules expressed on the surface of antigen-presenting cells. TCRαβ subunits are associated with the CD3 complex formed by the γ, δ, ε, and ζ subunits, which are invariable and ensure signal transduction. Here, we review how the expression and function of TCR complexes are orchestrated by several fine-tuned cellular processes that encompass (a) synthesis of the subunits and their correct assembly and expression at the plasma membrane as a single functional complex, (b) TCR membrane localization and dynamics at the plasma membrane and in endosomal compartments, (c) TCR signal transduction leading to T cell activation, and (d) TCR degradation. These processes balance each other to ensure efficient T cell responses to a variety of antigenic stimuli while preventing autoimmunity.
Published: 2018

9. Paclitaxel Response Can Be Predicted With Interpretable Multi-Variate Classifiers Exploiting DNA-Methylation and miRNA Data

Author: Borazjani, Nassim, Sepehri, Saghi, Behzadi, Maryam, Jarrahpour, Aliasghar, Rad, Javad Ameri, Sasanipour, Maryam, Mohkam, Milad, Ghasemi, Younes, Akbarizadeh, Amin Reza, Digiorgio, Carole, Brunel, Jean Michel, Ghanbari, Mohammad Mehdi, Batta, Gyula, Turos, Edward, Shiraz University (Shiraz University ), Ardabil University of Medical Sciences, Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Islamic Azad University, University of Debrecen Egyetem [Debrecen], University of South Florida [Tampa] (USF), Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU), and University of South Florida (USF)
Subjects: 0301 basic medicine, MESH: Inflammation, Cytotoxicity, [SDV]Life Sciences [q-bio], β-Lactams, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, 0302 clinical medicine, MESH: Structure-Activity Relationship, biomarker discovery, MESH: RAW 264.7 Cells, MESH: Animals, MESH: Neoplasms, Biomarker discovery, Genetics (clinical), Original Research, MESH: Drug Screening Assays, Antitumor, Methylation, artificial intelligence, MESH: Anti-Inflammatory Agents, Non-Steroidal, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], tumor profiling, 3. Good health, Colon cancer, machine learning, Anticancer, Paclitaxel, CpG site, MESH: Cell Survival, 030220 oncology & carcinogenesis, precision oncology, DNA methylation, Molecular Medicine, MESH: Drug Evaluation, Preclinical, lcsh:QH426-470, MESH: Molecular Structure, Decision tree, MESH: Hep G2 Cells, [SDV.CAN]Life Sciences [q-bio]/Cancer, Computational biology, Biology, 03 medical and health sciences, Breast cancer, MESH: beta-Lactams, MESH: Cell Proliferation, microRNA, medicine, Genetics, MESH: Tumor Cells, Cultured, MESH: Mice, DABCO, MESH: Humans, medicine.disease, lcsh:Genetics, 030104 developmental biology, chemistry, MESH: Benzopyrans, MESH: Antineoplastic Agents, Anti-inflammatory, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: International audience; To address the problem of resistance to paclitaxel treatment, we have investigated to which extent is possible to predict Breast Cancer (BC) patient response to this drug. We carried out a large-scale tumor-based prediction analysis using data from the US National Cancer Institute's Genomic Data Commons. These data sets comprise the responses of BC patients to paclitaxel along with six molecular profiles of their tumors. We assessed 10 Machine Learning (ML) algorithms on each of these profiles and evaluated the resulting 60 classifiers on the same BC patients. DNA methylation and miRNA profiles were the most informative overall. In combination with these two profiles, ML algorithms selecting the smallest subset of molecular features generated the most predictive classifiers: a complexity-optimized XGBoost classifier based on CpG island methylation extracted a subset of molecular factors relevant to predict paclitaxel response (AUC = 0.74). A CpG site methylation-based Decision Tree (DT) combining only 2 of the 22,941 considered CpG sites (AUC = 0.89) and a miRNA expression-based DT employing just 4 of the 337 analyzed mature miRNAs (AUC = 0.72) reveal the molecular types associated to paclitaxel-sensitive and resistant BC tumors. A literature review shows that features selected by these three classifiers have been individually linked to the cytotoxic-drug sensitivities and prognosis of BC patients. Our work leads to several molecular signatures, unearthed from methylome and miRNome, able to anticipate to some extent which BC tumors respond or not to paclitaxel. These results may provide insights to optimize paclitaxel-therapies in clinical practice.
Published: 2019

10. Structure, regulation, and (patho-)physiological functions of the stress-induced protein kinase CK1 delta (CSNK1D)

Author: Joachim Bischof, Timo Burster, Pengfei Xu, Fabian Gärtner, Congxing Liu, Vasiliy A. Bakulev, Uwe Knippschild, Najma Rachidi, Chiara Ianes, Universitätsklinikum Ulm - University Hospital of Ulm, Nazarbayev University [Kazakhstan], Ural Federal University [Ekaterinburg] (UrFU), Parasitologie moléculaire et Signalisation / Molecular Parasitology and Signaling, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by a grant to Uwe Knippschild from the Deutsche Forschungsgemeinschaft (DFG) [grant number KN356/9-1], a grant to Joachim Bischof from the Else Kröner-Fresenius-Stiftung [grant number 2017_A142], and a grant to Najma Rachidi [grant number ANR-13-ISV3-0009]., ANR-13-ISV3-0009,TranSig,Trans-signalisation: Un nouveau mécanisme permettant à Leishmania d'éviter la réponse immunitaire de la cellule hôte par la sécrétion de protéines de signalisation(2013), and Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: 0301 basic medicine, p53, MESH: Neoplasm Proteins, MESH: Signal Transduction, [SDV]Life Sciences [q-bio], Hedgehog pathway, 0302 clinical medicine, Drug Delivery Systems, MESH: Structure-Activity Relationship, Neoplasms, MESH: Animals, MESH: Neoplasms, Phosphorylation, Cancer, Kinase, Wnt signaling pathway, General Medicine, Stress-induced kinase, Hedgehog signaling pathway, 3. Good health, Neoplasm Proteins, Isoenzymes, CSNK1D, 030220 oncology & carcinogenesis, Casein Kinase Idelta, MESH: Isoenzymes, Casein kinase 1, Small molecule inhibitor, Signal Transduction, Gene isoform, MESH: Drug Delivery Systems, [SDV.CAN]Life Sciences [q-bio]/Cancer, Computational biology, Biology, Article, 03 medical and health sciences, Structure-Activity Relationship, Genetics, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology, Stress-Induced Protein, MESH: Humans, Site-specific phosphorylation, 030104 developmental biology, MESH: Casein Kinase Idelta
Abstract: International audience; Members of the highly conserved pleiotropic CK1 family of serine/threonine-specific kinases are tightly regulated in the cell and play crucial regulatory roles in multiple cellular processes from protozoa to human. Since their dysregulation as well as mutations within their coding regions contribute to the development of various different pathologies, including cancer and neurodegenerative diseases, they have become interesting new drug targets within the last decade. However, to develop optimized CK1 isoform-specific therapeutics in personalized therapy concepts, a detailed knowledge of the regulation and functions of the different CK1 isoforms, their various splice variants and orthologs is mandatory. In this review we will focus on the stress-induced CK1 isoform delta (CK1δ), thereby addressing its regulation, physiological functions, the consequences of its deregulation for the development and progression of diseases, and its potential as therapeutic drug target.
Published: 2019

11. Conserved domains and structure prediction of human cytomegalovirus UL27 protein

Author: Gaël Champier, Anthony Couvreux, Laurène Trapes, Sébastien Cotin, Sébastien Hantz, François Denis, Sophie Alain, Serge Bouaziz, Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, Laboratoire d'informatique Fondamentale de Marseille - UMR 6166 (LIF), Université de la Méditerranée - Aix-Marseille 2-Université de Provence - Aix-Marseille 1-Centre National de la Recherche Scientifique (CNRS), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC), Bouaziz, Serge, Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Human cytomegalovirus, MESH: Sequence Analysis, DNA, viruses, Cytomegalovirus, MESH: Amino Acid Sequence, medicine.disease_cause, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Pharmacology (medical), Peptide sequence, Conserved Sequence, Mutation, MESH: Conserved Sequence, MESH: Drug Resistance, Viral, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], virus diseases, Maribavir, Cell biology, Phosphotransferases (Alcohol Group Acceptor), Infectious Diseases, MESH: Antiviral Agents, MESH: Cytomegalovirus, MESH: Mutation, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Sequence analysis, Molecular Sequence Data, MESH: Sequence Alignment, Sequence alignment, Biology, Antiviral Agents, Herpesviridae, Structure-Activity Relationship, Drug Resistance, Viral, medicine, Humans, Amino Acid Sequence, Pharmacology, MESH: Ribonucleosides, MESH: Humans, MESH: Molecular Sequence Data, Sequence Analysis, DNA, medicine.disease, Virology, MESH: Phosphotransferases (Alcohol Group Acceptor), MESH: DNA, Viral, chemistry, DNA, Viral, Benzimidazoles, Ribonucleosides, MESH: Benzimidazoles, Sequence Alignment, DNA
Abstract: Background The human cytomegalovirus (HCMV) nuclear UL27 protein (pUL27) could be involved at the stage of nuclear egress. Maribavir is a new anti-HCMV drug that targets nuclear egress through direct inhibition of the HCMV serine–threonine kinase, UL97 protein (pUL97). Because maribavir- resistance-related mutations are observed in both proteins, pUL27 is thought to interfere with pUL97 activity; however, its mechanism of action remains unclear. Methods As there is no available crystal structure for pUL27 or any known structures of its homologous proteins, we attempted to identify pUL27 functional domains by sequence analysis, identification of conserved domains, structure prediction and matching with previously known maribavir resistance mutations. Results The UL27 sequence analysis of 20 HCMV wild-type strains and 8 ganciclovir-resistant HCMV strains allowed us to describe four conserved domains, to localize the putative phosphorylation sites and to identify protein–protein interface domains, suggesting that pUL27 could interact with either pUL97 or itself. Conclusions Although the function of pUL27 is still unknown in the HCMV replication cycle, our approach identified target domains that appeared to be essential to the function of pUL27. This work provides a better understanding on the relative importance of each pUL27 mutation and could form the basis of later comparison analyses, when a three-dimensional structure of a pUL27 homologue will be available.
Published: 2019

12. The 3-O-(3',3'-dimethylsuccinyl) derivative of betulinic acid (DSB) inhibits the assembly of virus-like particles in HIV-1 Gag precursor-expressing cells

Author: Sandrina Dafonseca, Armand Blommaert, Pascale CORIC, Saw See Hong, Serge Bouaziz, Pierre Boulanger, Centre de recherche, CHU Montréal, Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Infections Virales et Pathologie Comparée - UMR 754 (IVPC), Institut National de la Recherche Agronomique (INRA)-École pratique des hautes études (EPHE)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Rétrovirus et Pathologie Comparée (RPC), Université de Lyon-Université de Lyon-Ecole Nationale Vétérinaire de Lyon (ENVL), Bouaziz, Serge, Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Claude Bernard Lyon 1 (UCBL), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)
Subjects: Models, Molecular, MESH: Protein Precursors, MESH: Virus Assembly, Anti-HIV Agents, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MESH: Succinates, Spodoptera, Cell Line, MESH: Dose-Response Relationship, Drug, MESH: HIV-1, Structure-Activity Relationship, MESH: Protein Structure, Tertiary, MESH: Structure-Activity Relationship, Animals, MESH: Animals, Protein Precursors, MESH: Anti-HIV Agents, MESH: Spodoptera, Dose-Response Relationship, Drug, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Virus Assembly, MESH: Triterpenes, Succinates, Triterpenes, Protein Structure, Tertiary, MESH: Cell Line, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], MESH: Baculoviridae, HIV-1, Baculoviridae, MESH: Models, Molecular
Abstract: The 3-O-(3',3'-dimethylsuccinyl) derivative of betulinic acid (DSB) blocks HIV-1 maturation by interfering with viral protease (PR) at the capsid (CA)-SP1 cleavage site, a crucial region in HIV-1 morphogenesis.We analysed the effect of DSB on the assembly of HIV-1 Gag precursor (Pr55Gag(HIV)) into membrane-enveloped virus-like particles (VLP) in baculovirus-infected cells expressing Pr55Gag(HIV), in a cellular context devoid of viral PR.DSB showed a dose-dependent negative effect on VLP assembly, with an IC50 approximately 10 microM. The DSB inhibitory effect was p6-independent and was also observed for intracellular assembly of non-N-myristoylated Gag core-like particles. HIV-1 VLP assembled in the presence of DSB exhibited a lower stability of their inner cores upon membrane delipidation compared with control VLP, suggesting weaker Gag-Gag interactions. DSB also inhibited the assembly of simian immunodeficiency virus SLVmac251 VLP, although with a twofold lower efficacy (IC50 approximately 20 microM). No detectable inhibitory activity was observed for murine leukaemia virus (MLV) VLP; however, fusion of the SP1-NC-p6 domains from HIV-1 to the matrix (MA)-CA domains from MLV conferred DSB sensitivity to the chimaeric Gag precursor Pr72Gag(MLV-HIV) (IC50 = 30 microM). This observation suggested that the main DSB target on Pr55Gag was the SP1 domain, but the higher degree of DSB resistance for Pr72Gag(MLV-HIV) compared with Pr55Gag(HIV) implied that other upstream Gag region(s) might contribute to DSB reactivity.Sequence alignment and three-dimensional modelling by homology of the CA-SP1-NC junction in HIV-1, SLVmac251 and Pr72Gag(MLV-HIV) suggested that a higher hydrophilic character of the CA region immediately upstream to the HIV-1 CA-SP1 junction, as occurred in Pr72Gag(MLV-HIV), correlated with a lower DSB sensitivity.
Published: 2019

13. Substrate binding mode and catalytic mechanism of human heparan sulfate d-glucuronyl C5 epimerase

Author: Emeline Richard, Véronique Roig-Zamboni, Hugues Lortat-Jacob, Firas Fadel, Jérôme Hénault, Adeline Goulet, Christine Le Narvor, Yves Bourne, David Bonnaffé, Vincent Delauzun, C. Debarnot, Yoan R. Monneau, Bernard Priem, Romain R. Vivès, Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Institut de Chimie Moléculaire et des Matériaux d'Orsay (ICMMO), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Centre de Recherches sur les Macromolécules Végétales (CERMAV), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Institut de Chimie du CNRS (INC)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'ingénierie des systèmes macromoléculaires (LISM), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU), ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), ANR-10-LABX-0049,GRAL,Grenoble Alliance for Integrated Structural Cell Biology(2010), ANR-11-IDEX-0003,IPS,Idex Paris-Saclay(2011), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), LCOM eG2M, Inst Chim Mol & Mat Orsay, LabEx LERMIT,UMR 8182, Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherches sur les Macromolécules Végétales (CERMAV ), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche en neurobiologie - neurophysiologie de Marseille (CRN2M), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Bourne, Yves, Infrastructure Française pour la Biologie Structurale Intégrée - - FRISBI2010 - ANR-10-INBS-0005 - INBS - VALID, Grenoble Alliance for Integrated Structural Cell Biology - - GRAL2010 - ANR-10-LABX-0049 - LABX - VALID, and Idex Paris-Saclay - - IPS2011 - ANR-11-IDEX-0003 - IDEX - VALID
Subjects: MESH: Heparin, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Oligosaccharides, Isomerase, Uronic acid, catalytic mechanism, Crystallography, X-Ray, 01 natural sciences, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Substrate Specificity, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Glucuronic Acid, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Trisaccharide, [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], chemistry.chemical_classification, [SDV.MHEP.ME] Life Sciences [q-bio]/Human health and pathology/Emerging diseases, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, 0303 health sciences, [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, Multidisciplinary, [SDV.BIBS] Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Nuclear magnetic resonance spectroscopy, Heparan sulfate, Biological Sciences, MESH: Carbohydrate Epimerases, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], MESH: Glucuronic Acid, [SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, MESH: HEK293 Cells, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, heparan sulfate, substrate distortion, Stereochemistry, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Catalysis, Structure-Activity Relationship, 03 medical and health sciences, Humans, Binding site, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.IMM.II] Life Sciences [q-bio]/Immunology/Innate immunity, 030304 developmental biology, X-ray crystallography, Binding Sites, MESH: Humans, Heparin, 010405 organic chemistry, Mandelate racemase, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Active site, MESH: Catalysis, MESH: Crystallography, X-Ray, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 0104 chemical sciences, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, HEK293 Cells, chemistry, MESH: Binding Sites, C5 epimerization, biology.protein, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, MESH: Substrate Specificity, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Carbohydrate Epimerases, MESH: Oligosaccharides
Abstract: Heparan sulfate (HS) is a linear, complex polysaccharide that modulates the biological activities of proteins through binding sites made by a series of Golgi-localized enzymes. Of these, glucuronyl C5-epimerase (Glce) catalyzes C5-epimerization of the HS component, d -glucuronic acid (GlcA), into l -iduronic acid (IdoA), which provides internal flexibility to the polymer and forges protein-binding sites to ensure polymer function. Here we report crystal structures of human Glce in the unbound state and of an inactive mutant, as assessed by real-time NMR spectroscopy, bound with a (GlcA-GlcNS) n substrate or a (IdoA-GlcNS) n product. Deep infiltration of the oligosaccharides into the active site cleft imposes a sharp kink within the central GlcNS-GlcA/IdoA-GlcNS trisaccharide motif. An extensive network of specific interactions illustrates the absolute requirement of N -sulfate groups vicinal to the epimerization site for substrate binding. At the epimerization site, the GlcA/IdoA rings are highly constrained in two closely related boat conformations, highlighting ring-puckering signatures during catalysis. The structure-based mechanism involves the two invariant acid/base residues, Glu499 and Tyr578, poised on each side of the target uronic acid residue, thus allowing reversible abstraction and readdition of a proton at the C5 position through a neutral enol intermediate, reminiscent of mandelate racemase. These structures also shed light on a convergent mechanism of action between HS epimerases and lyases and provide molecular frameworks for the chemoenzymatic synthesis of heparin or HS analogs.
Published: 2019

14. New insight into structure-activity of furan-based salicylate synthase (MbtI) inhibitors as potential antitubercular agents

Author: Chiarelli, Laurent, Mori, Matteo, Beretta, Giangiacomo, Gelain, Arianna, Pini, Elena, Sammartino, Josè Camilla, Stelitano, Giovanni, Barlocco, Daniela, Costantino, Luca, Lapillo, Margherita, Poli, Giulio, Caligiuri, Isabella, Rizzolio, Flavio, Bellinzoni, Marco, Tuccinardi, Tiziano, Villa, Stefania, Meneghetti, Fiorella, University of Pavia, Università degli Studi di Milano [Milano] (UNIMI), University of Modena and Reggio Emilia, Partenaires INRAE, University of Pisa - Università di Pisa, National Cancer Institute of Aviano, University of Ca’ Foscari [Venice, Italy], Microbiologie structurale - Structural Microbiology (Microb. Struc. (UMR_3528 / U-Pasteur_5)), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), This work was funded by University of Milan (Linea B), and the Italian Ministry of Education, University and Research (MIUR): Dipartimenti di Eccellenza Programme (2018–2022) - Dept. of Biology and Biotechnology 'L. Spallanzani', University of Pavia., Università degli Studi di Pavia = University of Pavia (UNIPV), Università degli Studi di Milano = University of Milan (UNIMI), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)
Subjects: MESH: Mycobacterium bovis, drug design, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Short Communication, mycobactins, Antitubercular Agents, Lyases, Settore BIO/11 - Biologia Molecolare, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Microbial Sensitivity Tests, Structure-Activity Relationship, MESH: Structure-Activity Relationship, MESH: Molecular Docking Simulation, [CHIM.CRIS]Chemical Sciences/Cristallography, antimycobacterial agent, Tuberculosis, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Enzyme Inhibitors, Furans, molecular modelling, siderophores, Binding Sites, Molecular Docking Simulation, Mycobacterium bovis, Pharmacology, Drug Discovery3003 Pharmaceutical Science, MESH: Microbial Sensitivity Tests, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], lcsh:RM1-950, MESH: Furans, MESH: Antitubercular Agents, MESH: Lyases, lcsh:Therapeutics. Pharmacology, MESH: Binding Sites, MESH: Enzyme Inhibitors, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM]
Abstract: Starting from the analysis of the hypothetical binding mode of our previous furan-based hit (I), we successfully achieved our objective to replace the nitro moiety, leading to the disclosure of a new lead exhibiting a strong activity against MbtI. Our best candidate 1 h displayed a Ki of 8.8 µM and its antimycobacterial activity (MIC99 = 250 µM) is conceivably related to mycobactin biosynthesis inhibition. These results support the hypothesis that 5-phenylfuran-2-carboxylic derivatives are a promising class of MbtI inhibitors., GRAPHICAL ABSTRACT
Published: 2019

15. Expression and purification of recombinant extracellular sulfatase HSulf-2 allows deciphering of enzyme sub-domain coordinated role for the binding and 6-O-desulfation of heparan sulfate

Author: Amal Seffouh, Hugues Lortat-Jacob, Zahra el Oula Hassoun, Rana El Masri, Jean-Pierre Andrieu, Romain R. Vivès, Evelyne Gout, Olga Makshakova, Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Kazan Institute of Biochemistry and Biophysics, Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and ANR-17-CE11-0040,SULFatAS,Structure et activités des sulfatases extracellulaires SULF(2017)
Subjects: Models, Molecular, Mutant, MESH: Catalytic Domain, Substrate Specificity, law.invention, MESH: Recombinant Proteins, chemistry.chemical_compound, MESH: Structure-Activity Relationship, law, Catalytic Domain, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, 0303 health sciences, Sulfatase, MESH: Hydrophobic and Hydrophilic Interactions, 030302 biochemistry & molecular biology, Extracellular matrix, Heparan sulfate, Recombinant Proteins, Cell biology, Structure–function relationships, MESH: HEK293 Cells, Recombinant DNA, Molecular Medicine, Sulfatases, Sulfotransferases, Hydrophobic and Hydrophilic Interactions, MESH: Models, Molecular, Protein Binding, Glycocalyx, Cell Line, Structure-Activity Relationship, 03 medical and health sciences, Cellular and Molecular Neuroscience, MESH: Heparitin Sulfate, Extracellular, Humans, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Binding site, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Molecular Biology, Pharmacology, Binding Sites, MESH: Humans, Heparin, HEK 293 cells, Cell Biology, MESH: Sulfotransferases, MESH: Cell Line, HEK293 Cells, Enzyme, Glycosaminoglycan, MESH: Binding Sites, chemistry, MESH: Substrate Specificity, Heparitin Sulfate
Abstract: International audience; Through their ability to edit 6-O-sulfation pattern of Heparan sulfate (HS) polysaccharides, Sulf extracellular endosulfatases have emerged as critical regulators of many biological processes, including tumor progression. However, study of Sulfs remains extremely intricate and progress in characterizing their functional and structural features has been hampered by limited access to recombinant enzyme. In this study, we unlock this critical bottleneck, by reporting an efficient expression and purification system of recombinant HSulf-2 in mammalian HEK293 cells. This novel source of enzyme enabled us to investigate the way the enzyme domain organization dictates its functional properties. By generating mutants, we confirmed previous studies that HSulf-2 catalytic (CAT) domain was sufficient to elicit arylsulfatase activity and that its hydrophilic (HD) domain was necessary for the enzyme 6-O-endosulfatase activity. However, we demonstrated for the first time that high-affinity binding of HS substrates occurred through the coordinated action of both domains, and we identified and characterized 2 novel HS binding sites within the CAT domain. Altogether, our findings contribute to better understand the molecular mechanism governing HSulf-2 substrate recognition and processing. Furthermore, access to purified recombinant protein opens new perspectives for the resolution of HSulf structure and molecular features, as well as for the development of Sulf-specific inhibitors.
Published: 2019

16. Overview of the Structure–Function Relationships of Mannose-Specific Lectins from Plants, Algae and Fungi

Author: Pierre Rougé, Els J.M. Van Damme, Yves Bourne, Annick Barre, Pharmacochimie et Biologie pour le Développement (PHARMA-DEV), Institut de Recherche pour le Développement (IRD)-Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Architecture et fonction des macromolécules biologiques (AFMB), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Faculty of Bioscience Engineering [Ghent], Universiteit Gent = Ghent University (UGENT), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Institut de Recherche pour le Développement (IRD), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA), Universiteit Gent = Ghent University [Belgium] (UGENT), Surfaces Cellulaires et Signalisation chez les Végétaux (SCSV), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Laboratory of Biochemistry and Glycobiology, Chent University, and Institut de pharmacologie et de biologie structurale (IPBS)
Subjects: Models, Molecular, 0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Protein Conformation, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Plants, AMINO-ACID-SEQUENCE, Oligosaccharides, Mannose, plant, Review, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, ENVELOPE GLYCOPROTEIN GP120, lcsh:Chemistry, GLYCAN-SPECIFIC LECTIN, chemistry.chemical_compound, MESH: Protein Conformation, MESH: Structure-Activity Relationship, PINELLIA-TERNATA AGGLUTININ, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, CANAVALIA-BRASILIENSIS LECTIN, use as tools, lcsh:QH301-705.5, Peptide sequence, Spectroscopy, CARBOHYDRATE-BINDING, [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Chemistry, Structure function, General Medicine, Plants, Envelope glycoprotein GP120, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Computer Science Applications, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], JACALIN-RELATED, Biochemistry, mannose-binding specificity, MESH: Mannose, HUMAN-IMMUNODEFICIENCY-VIRUS, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, LECTINS, MESH: Models, Molecular, Protein Binding, MESH: Fungi, Glycan, Catalysis, Inorganic Chemistry, Structure-Activity Relationship, 03 medical and health sciences, Algae, MESH: Mannose-Binding Lectins, MESH: Protein Binding, structure, Physical and Theoretical Chemistry, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], AGENTS, ANTI-HIV ACTIVITY, Molecular Biology, function, Binding Sites, 030102 biochemistry & molecular biology, Organic Chemistry, Biology and Life Sciences, Lectin, biology.organism_classification, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Mannose-Binding Lectins, 030104 developmental biology, lcsh:Biology (General), lcsh:QD1-999, MESH: Binding Sites, MANNOSE/GLUCOSE-SPECIFIC LECTIN, biology.protein, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, lectin, fungi, MESH: Oligosaccharides, Function (biology)
Abstract: International audience; To date, a number of mannose-binding lectins have been isolated and characterized from plants and fungi. These proteins are composed of different structural scaffold structures which harbor a single or multiple carbohydrate-binding sites involved in the specific recognition of mannose-containing glycans. Generally, the mannose-binding site consists of a small, central, carbohydrate-binding pocket responsible for the "broad sugar-binding specificity" toward a single mannose molecule, surrounded by a more extended binding area responsible for the specific recognition of larger mannose-containing N-glycan chains. Accordingly, the mannose-binding specificity of the so-called mannose-binding lectins towards complex mannose-containing N-glycans depends largely on the topography of their mannose-binding site(s). This structure⁻function relationship introduces a high degree of specificity in the apparently homogeneous group of mannose-binding lectins, with respect to the specific recognition of high-mannose and complex N-glycans. Because of the high specificity towards mannose these lectins are valuable tools for deciphering and characterizing the complex mannose-containing glycans that decorate both normal and transformed cells, e.g., the altered high-mannose N-glycans that often occur at the surface of various cancer cells.
Published: 2019

17. Identification of novel quinazoline derivatives as potent antiplasmodial agents

Author: Johan Schultz, Dany Pechalrieu, Sergio Valente, Anne Bouchut, Tina S. Skinner-Adams, Giancarlo Fabrizi, Paola B. Arimondo, Urban Hoglund, Katherine T. Andrews, Roberta Mazzone, Sophia Lafitte, Antonello Mai, Alessia Lucidi, Christine Pierrot, Ming Jang Chua, Jamal Khalife, Dante Rotili, Institut Pasteur de Lille, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome] [UNIROMA], Pharmacochimie de la Régulation Epigénétique du Cancer [ETaC], Réseau International des Instituts Pasteur (RIIP), Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Kancera AB [Sweden], Adlego Biomedical AB [Sweden], Pharmacochimie de la Régulation Epigénétique du Cancer (ETaC), PIERRE FABRE-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), GRIFFITH UNIVERSITY AUS, Partenaires IRSTEA, Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA), This work was supported by the Australian National Health and Medical Research Council (APP1074016 to K.T.A.), Italian PRIN 2016 (prot. 20152TE5PK to A.M.), AIRC 2016 (n. 19162 to A.M.), NIH (n. R01GM114306 to A.M.), Progetto Ateneo Sapienza 2017 (to D.R.), the A-ParaDDisE program funded under the European Union's Seventh Framework Programme (grant agreement no. 602080 to A.B., C.P., S.L., A.M., D.R., S.V., J.S., U.H., K.T.A. and J.K.) and Griffith University (GUIPRS and GUPRS scholarships to M.J.C.). We thank Mary Clarke (Griffith University) for technical assistance. We also thank the BioImaging Center Lille Facility for access to the BD FACSCanto II cytometer., European Project: 602080,EC:FP7:HEALTH,FP7-HEALTH-2013-INNOVATION-1,A-PARADDISE(2014), Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome] (UNIROMA), Griffith University [Brisbane], Centre d’Infection et d’Immunité de Lille (CIIL) - INSERM U1019 - UMR 9017 (CIIL), Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Università degli Studi di Roma 'La Sapienza' [Rome], and PIERRE FABRE-Centre National de la Recherche Scientifique (CNRS)
Subjects: Plasmodium berghei, Molecular Conformation, Pharmacology, MESH: Parasitic Sensitivity Tests, 01 natural sciences, Antimalarial agents, MESH: Dose-Response Relationship, Drug, Mice, MESH: Structure-Activity Relationship, Parasitic Sensitivity Tests, PK studies, Histone deacetylase inhibitors, Drug Discovery, P. berghei mouse model, MESH: Animals, Antimalarial Agent, DNA methyltransferase inhibitors, Artemisinin, MESH: Plasmodium falciparum, media_common, 0303 health sciences, Mice, Inbred BALB C, biology, Chemistry, General Medicine, 3. Good health, MESH: Quinazolines, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, medicine.drug, Drug, antimalarial agents, histone deacetylase inhibitors, media_common.quotation_subject, Plasmodium falciparum, MESH: Mice, Inbred BALB C, Context (language use), 03 medical and health sciences, Antimalarials, Structure-Activity Relationship, In vivo, parasitic diseases, medicine, MESH: Plasmodium berghei, [CHIM]Chemical Sciences, Animals, Humans, MESH: Mice, 030304 developmental biology, MESH: Molecular Conformation, MESH: Humans, Dose-Response Relationship, Drug, 010405 organic chemistry, Organic Chemistry, Fibroblasts, biology.organism_classification, medicine.disease, MESH: Antimalarials, 0104 chemical sciences, MESH: Fibroblasts, Quinazolines, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Malaria
Abstract: International audience; Despite the recent reductions in the global burden of malaria, this disease remains a devastating cause of death in tropical and subtropical regions. As there is no broadly effective vaccine for malaria, prevention and treatment still rely on chemotherapy. Unfortunately, emerging resistance to the gold standard artemisinin combination therapies means that new drugs with novel modes of action are urgently needed. In this context, Plasmodium histone modifying enzymes have emerged as potential drug targets, prompting us to develop and optimize compounds directed against such epigenetic targets. A panel of 51 compounds designed to target different epigenetic enzymes were screened for activity against Plasmodium falciparum parasites. Based on in vitro activity against drug susceptible and drug-resistant P. falciparum lines, selectivity index criterion and favorable pharmacokinetic properties, four compounds, one HDAC inhibitor (1) and three DNMT inhibitors (37, 43 and 45), were selected for preclinical studies in a mouse model of malaria. In vivo data showed that 37, 43 and 45 exhibited oral efficacy in the mouse model of Plasmodium berghei infection. These compounds represent promising starting points for the development of novel antimalarial drugs.
Published: 2019

18. 8-Thioalkyl-adenosine derivatives inhibit Listeria monocytogenes NAD kinase through a novel binding mode

Author: Julie Paoletti, Gilles Labesse, Olivier Dussurget, Marie Anne Nahori, Liliane Assairi, Sylvie Pochet, Muriel Gelin, Valérie Huteau, Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), U759 Unité Imagerie intégrative, Institut Curie, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Biochimie Structurale [Montpellier] (CBS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS), Université de Montpellier (UM), Interactions Bactéries-Cellules (UIBC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), Université Sorbonne Paris Cité (USPC), Agence Nationale de la Recherche [ANR-06-BLAN-0324, ANR-11-EMM-0019], French Infrastructure for Integrated Structural Biology (FRISBI) [ANR-10-INSB-05-01], ANR [ANR-11-EMM-0019], ANR-10-INBS-05-01/10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), ANR-11-EMMA-0019,ANAKIN (Anti-NAd-Kinase),Nouveaux composés antibactériens ciblant la NAD kinase(2011), ANR-06-BLAN-0324,Gram(+) NADK,Rational design and in situ selection of drug targetting NAD kinases from Gram(+) bacterial pathogens(2006), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: 0301 basic medicine, Adenosine, MESH: Adenosine/chemistry, Protein Conformation, Ribose, [SDV]Life Sciences [q-bio], MESH: Staphylococcus aureus/enzymology, MESH: Amino Acid Sequence, MESH: Ribose/chemistry, Nicotinamide adenine dinucleotide, 01 natural sciences, MESH: Adenosine/pharmacology, chemistry.chemical_compound, MESH: Protein Conformation, MESH: Structure-Activity Relationship, Drug Discovery, Transferase, Enzyme Inhibitors, MESH: Listeria monocytogenes/enzymology, biology, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Kinase, General Medicine, MESH: Phosphotransferases (Alcohol Group Acceptor)/metabolism, Molecular Docking Simulation, 010101 applied mathematics, Phosphotransferases (Alcohol Group Acceptor), Biochemistry, Phosphorylation, Nicotinamide adenine dinucleotide phosphate, Protein Binding, medicine.drug, Staphylococcus aureus, Inhibitor, MESH: Phosphotransferases (Alcohol Group Acceptor)/chemistry, Stereochemistry, Cofactor, Structure-Activity Relationship, 03 medical and health sciences, MESH: Adenosine/metabolism, Adenosine analogue, MESH: Molecular Docking Simulation, medicine, Humans, MESH: Protein Binding, Amino Acid Sequence, MESH: Enzyme Inhibitors/chemistry, 0101 mathematics, NAD kinase, X-ray crystallography, Pharmacology, MESH: Humans, MESH: Enzyme Inhibitors/pharmacology, Organic Chemistry, MESH: Enzyme Inhibitors/metabolism, Listeria monocytogenes, Antibacterial, 030104 developmental biology, chemistry, biology.protein, NAD+ kinase, MESH: Phosphotransferases (Alcohol Group Acceptor)/antagonists & inhibitors
Abstract: Increased resistance of pathogens to existing antibiotics necessitates the search for novel targets to develop potent antimicrobials. Biosynthetic pathways of several cofactors important for bacterial growth, such as nicotinamide adenine dinucleotide phosphate (NADP), have been proposed as a promising source of antibiotic targets. Nicotinamide adenine dinucleotide kinases (NADK; EC 2.7.1.23) are attractive for inhibitor development, since they catalyze the phosphorylation of NAD to NADP, which is an essential step of NADP metabolism. We previously synthesized diadenosine derivatives that inhibited NADK from two human pathogens, Listeria monocytogenes and Staphylococcus aureus, in the micromolar range. They behave as NAD mimics with the 5',5'-diphosphate group substituted by a 8,5' thioglycolic bridge. In an attempt to improve inhibitory potency, we designed new NAD mimics based on a single adenosine moiety harboring a larger derivatization attached to the C8 position and a small group at the 5' position. Here we report the synthesis of a series of 8-thioalkyl-adenosine derivatives containing various aryl and heteroaryl moieties and their evaluation as inhibitors of L. monocytogenes NADK1, S. aureus NADK and their human counterpart. Novel, sub-micromolar inhibitors of LmNADK1 were identified. Surprisingly, most LmNADK1 inhibitors demonstrated a high selectivity index against the close staphylococcal ortholog and the human NADK. Structural characterization of enzyme-inhibitor complexes revealed the original binding mode of these novel NAD mimics.
Published: 2016

19. Guttiferone A Aggregates Modulate Silent Information Regulator 1 (SIRT1) Activity

Author: Cottet, Kévin, Xu, Bin, Coric, Pascale, Bouaziz, Serge, Michel, Michel, Vidal, Michel, Lallemand, Marie-Christine, Broussy, Sylvain, Michel, Sylvie, Yangtze Delta Region Institute of Tsinghua University [Zhejiang], Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Institute of cardiometabolism and nutrition (ICAN), Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Equipe Pharmacognosie (UMR 8638), Chimie Organique, Médicinale et Extractive et Toxicologie Expérimentale (COMETE - UMR 8638), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université Paris Descartes - Paris 5 (UPD5), Laboratoire de chimie de coordination (LCC), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Mécanique et Technologie (LMT), École normale supérieure - Cachan (ENS Cachan)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases (ICAN), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), and Bouaziz, Serge
Subjects: Models, Molecular, 0301 basic medicine, MESH: Benzophenones, Magnetic Resonance Spectroscopy, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MESH: Biological Products, MESH: Molecular Structure, Regulator, MESH: Dynamic Light Scattering, MESH: Dose-Response Relationship, Drug, Benzophenones, Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Sirtuin 1, Clusiaceae, Drug Discovery, Humans, Structure–activity relationship, MESH: Sirtuin 1, Enzyme Inhibitors, chemistry.chemical_classification, Biological Products, MESH: Humans, Natural product, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Dose-Response Relationship, Drug, Molecular Structure, MESH: Magnetic Resonance Spectroscopy, Nuclear magnetic resonance spectroscopy, Dynamic Light Scattering, In vitro, 3. Good health, MESH: Clusiaceae, Hyperforin, 030104 developmental biology, Enzyme, chemistry, Biochemistry, MESH: Enzyme Inhibitors, Biological target, Molecular Medicine, MESH: Models, Molecular
Abstract: International audience; Natural products guttiferone A, hyperforin, and aristoforin were able to inhibit or increase SIRT1 catalytic activity, depending on protein concentration and presence of detergent. On the basis of NMR data for guttiferone A, we demonstrated that the aggregation state of the natural product played a crucial role for its interaction with the enzyme. These results are useful to interpret future in vitro structure-activity relationship studies on these natural products in the quest of their biological target(s).
Published: 2016

20. Structure–function analysis of the extracellular domain of the pneumococcal cell division site positioning protein MapZ

Author: Sylvie Manuse, Cédric Laguri, Jean-Pierre Simorre, Michael S. VanNieuwenhze, Jean-Pierre Lavergne, Catherine M. Bougault, Nicolas L. Jean, Mégane Guinot, Christophe Grangeasse, Microbiologie moléculaire et biochimie structurale / Molecular Microbiology and Structural Biochemistry (MMSB), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Department of Chemistry, Indiana University, Indiana University [Bloomington], Indiana University System-Indiana University System, PLATIM and Protein Sciences platforms of SFR Biosciences Gerland-Lyon Sud (UMS344/US8), NMR and isotope labelling platforms of the Grenoble Instruct Center (ISBG, UMS 3518 CNRS-CEA-UJF-EMBL), ANR-12-BSV3-0008,PiBaKi,Rôle cellulaire et mode d'action de deux protéine-kinases centrales chez les bactéries(2012), ANR-15-CE32-0001,Map-CellDiv,MapZ: caractérisation d'un nouveau mécanisme de régulation de la division cellulaire bactérienne(2015), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Subjects: MESH: Cytokinesis, 0301 basic medicine, Models, Molecular, MESH: Cytoskeletal Proteins, Cell division, Protein Conformation, Science, Protein domain, General Physics and Astronomy, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, Structure-Activity Relationship, MESH: Protein Conformation, MESH: Structure-Activity Relationship, Protein structure, Bacterial Proteins, Protein Domains, FtsZ, Cytoskeleton, MESH: Bacterial Proteins, Cytokinesis, MESH: Gene Expression Regulation, Bacterial, Multidisciplinary, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], biology, General Chemistry, Gene Expression Regulation, Bacterial, Cell cycle, Cell biology, Cytoskeletal Proteins, 030104 developmental biology, Streptococcus pneumoniae, Biochemistry, biology.protein, MESH: Cell Division, MESH: Protein Domains, MESH: Models, Molecular, MESH: Streptococcus pneumoniae, Cell Division, Cell division site
Abstract: Accurate placement of the bacterial division site is a prerequisite for the generation of two viable and identical daughter cells. In Streptococcus pneumoniae, the positive regulatory mechanism involving the membrane protein MapZ positions precisely the conserved cell division protein FtsZ at the cell centre. Here we characterize the structure of the extracellular domain of MapZ and show that it displays a bi-modular structure composed of two subdomains separated by a flexible serine-rich linker. We further demonstrate in vivo that the N-terminal subdomain serves as a pedestal for the C-terminal subdomain, which determines the ability of MapZ to mark the division site. The C-terminal subdomain displays a patch of conserved amino acids and we show that this patch defines a structural motif crucial for MapZ function. Altogether, this structure–function analysis of MapZ provides the first molecular characterization of a positive regulatory process of bacterial cell division., Placement of the bacterial division site is crucial for the creation of identical daughter cells. Here, the authors solve the structure of the MapZ protein, which helps to position the cell division protein FtsZ at the cell centre, and further analyse the function of the protein in vivo.
Published: 2016

21. Tetramerization and interdomain flexibility of the replication initiation controller YabA enables simultaneous binding to multiple partners

Author: Liza Felicori, Magali Ventroux, Transito Garcia-Garcia, Laurent Terradot, Franck Molina, Anthony J. Wilkinson, Mark J. Fogg, Philippe Noirot, Katie H. Jameson, Alexandre Bazin, Mickaël V. Cherrier, Marie Francoise Noirot-Gros, Pierre Roblin, Sysdiag CNRS Bio-Rad UMR 3145, Cap Delta/Parc Euromédecine, Centre National de la Recherche Scientifique (CNRS), Département Caractérisation et Elaboration des Produits Issus de l'Agriculture (CEPIA), Institut National de la Recherche Agronomique (INRA), York Structural Biology Laboratory, Department of Chemistry, University of York [York, UK], MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Institut de biologie structurale (IBS - UMR 5075), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Informatique, de Modélisation et d'Optimisation des Systèmes (LIMOS), Ecole Nationale Supérieure des Mines de St Etienne-Université Clermont Auvergne (UCA)-Centre National de la Recherche Scientifique (CNRS), Sysdiag-Modélisation et Ingénierie des Systèmes Complexes Biologiques pour le Diagnostic (SysDiag ), BIO-RAD-Centre National de la Recherche Scientifique (CNRS), Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Ecole Nationale Supérieure des Mines de St Etienne-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS), European integrated project, BaSysBio, Biotechnology and Biological Sciences Research Council, UK, EU-Marie Curie Project AMBER FP7-People [317338], CIBLE program from the region Rhones-alpes, MICALIS INRA, Felicori, Liza, Jameson, Katie H., Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), and Microbiologie moléculaire et biochimie structurale / Molecular Microbiology and Structural Biochemistry (MMSB)
Subjects: Models, Molecular, 0301 basic medicine, Protein Conformation, Amino Acid Motifs, Intracellular Space, dnaN, MESH: DNA Replication, Plasma protein binding, MESH: Amino Acid Sequence, MESH: Zinc, MESH: Amino Acid Motifs, Protein structure, MESH: Structure-Activity Relationship, MESH: Protein Conformation, Structural Biology, Protein Interaction Mapping, MESH: Bacterial Proteins, Genetics, DNA clamp, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MESH: Protein Multimerization, DNA-Binding Proteins, Protein Transport, Zinc, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], MESH: Intracellular Space, MESH: Models, Molecular, Bacillus subtilis, Protein Binding, DNA Replication, MESH: Protein Transport, MESH: Mutation, Molecular Sequence Data, MESH: Sequence Alignment, Biology, Structure-Activity Relationship, 03 medical and health sciences, Bacterial Proteins, Position-Specific Scoring Matrices, MESH: Protein Binding, Protein Interaction Domains and Motifs, Amino Acid Sequence, Homology modeling, MESH: Protein Interaction Domains and Motifs, Binding Sites, MESH: Molecular Sequence Data, MESH: Protein Interaction Mapping, DNA replication, MESH: Position-Specific Scoring Matrices, MESH: Bacillus subtilis, MESH: Multiprotein Complexes, DnaA, 030104 developmental biology, MESH: Binding Sites, Replication Initiation, Multiprotein Complexes, Mutation, Biophysics, Protein Multimerization, Sequence Alignment, MESH: DNA-Binding Proteins
Abstract: International audience; YabA negatively regulates initiation of DNA replication in low-GC Gram-positive bacteria. The protein exerts its control through interactions with the initiator protein DnaA and the sliding clamp DnaN. Here, we combined X-ray crystallography, X-ray scattering (SAXS), modeling and biophysical approaches, with in vivo experimental data to gain insight into YabA function. The crystal structure of the N-terminal domain (NTD) of YabA solved at 2.7 Å resolution reveals an extended α-helix that contributes to an intermolecular four-helix bundle. Homology modeling and biochemical analysis indicates that the C-terminal domain (CTD) of YabA is a small Zn-binding domain. Multi-angle light scattering and SAXS demonstrate that YabA is a tetramer in which the CTDs are independent and connected to the N-terminal four-helix bundle via flexible linkers. While YabA can simultaneously interact with both DnaA and DnaN, we found that an isolated CTD can bind to either DnaA or DnaN, individually. Site-directed mutagenesis and yeast-two hybrid assays identified DnaA and DnaN binding sites on the YabA CTD that partially overlap and point to a mutually exclusive mode of interaction. Our study defines YabA as a novel structural hub and explains how the protein tetramer uses independent CTDs to bind multiple partners to orchestrate replication initiation in the bacterial cell.
Published: 2016

22. A Novel Class of Schistosoma mansoni Histone Deacetylase 8 (HDAC8) Inhibitors Identified by Structure-Based Virtual Screening and In Vitro Testing

Author: Raymond J. Pierce, Wolfgang Sippl, Julien Lancelot, Karin Schmidtkunz, Alokta Chakrabarti, Dina Robaa, Jelena Melesina, Tajith B. Shaik, Christophe Romier, Conrad V. Simoben, Manfred Jung, Srinivasaraghavan Kannan, Martin Marek, Romier, Christophe, Martin-Luther-Universität Halle Wittenberg (MLU), Albert-Ludwigs-Universität Freiburg, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), This work and the authors of this article received funding from the European Union’s Seventh Framework Programme for Research, Technological Development and Demonstration under Grant Agreements 241865 (SEtTReND) and 602080 (A-ParaDDisE). Further support was received by the Deutsche Forschungsgemeinschaft (Ju-295/13-1, SI-868/13-1). MM, TBS and CR are supported by institutional funds from the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé et de la Recherche Médicale (INSERM) and the Université de Strasbourg., The authors acknowledge the support and the use of resources of the French Infrastructure for Integrated Structural Biology FRISBI ANR-10-INBS-05 and of Instruct-ERIC. We wish to thank members of the ESRF-EMBL joint structural biology groups and the SOLEIL synchrotron for the use of their beamline facilities and for help during data collection. We are grateful to Pierre Legrand (SOLEIL) for his kind assistance for data processing, Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, and Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)
Subjects: 0301 basic medicine, Pyrrolidines, MESH: Helminth Proteins/metabolism, MESH: Histone Deacetylases/metabolism, MESH: Histone Deacetylases/genetics, Pharmaceutical Science, MESH: Protein Structure, Secondary, Gene Expression, Apoptosis, Crystallography, X-Ray, Hydroxamic Acids, Molecular Docking Simulation, MESH: Helminth Proteins/antagonists & inhibitors, MESH: Chelating Agents/chemical synthesis, Protein Structure, Secondary, Analytical Chemistry, 0302 clinical medicine, MESH: Structure-Activity Relationship, MESH: Hydroxamic Acids/chemical synthesis, Drug Discovery, MESH: Zinc/metabolism, MESH: Animals, [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], MESH: Histone Deacetylase Inhibitors/pharmacology, Chelating Agents, Anthelmintics, biology, Chemistry, MESH: Anthelmintics/pharmacology, Helminth Proteins, Schistosoma mansoni, epigenetics, crystal structure, docking, histone deacetylase (HDAC) inhibitors, schistosomiasis, virtual screening, MESH: Histone Deacetylases/chemistry, 3. Good health, MESH: Schistosoma mansoni/drug effects, MESH: Pyrrolidines/pharmacology, Zinc, Biochemistry, Chemistry (miscellaneous), Molecular Medicine, Protein Binding, MESH: Apoptosis/drug effects, MESH: Gene Expression, In silico, 030231 tropical medicine, MESH: Zinc/chemistry, Article, Histone Deacetylases, lcsh:QD241-441, 03 medical and health sciences, Structure-Activity Relationship, lcsh:Organic chemistry, MESH: Molecular Docking Simulation, Structure–activity relationship, MESH: Protein Binding, Animals, MESH: Helminth Proteins/genetics, Protein Interaction Domains and Motifs, Physical and Theoretical Chemistry, MESH: Histone Deacetylase Inhibitors/chemical synthesis, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Virtual screening, MESH: Protein Interaction Domains and Motifs, Binding Sites, Organic Chemistry, MESH: Hydroxamic Acids/pharmacology, HDAC8, MESH: Schistosoma mansoni/growth & development, Epigenome, biology.organism_classification, MESH: Crystallography, X-Ray, MESH: Anthelmintics/chemical synthesis, Histone Deacetylase Inhibitors, 030104 developmental biology, MESH: Binding Sites, Docking (molecular), MESH: Chelating Agents/pharmacology, MESH: Schistosoma mansoni/enzymology, MESH: Helminth Proteins/chemistry, MESH: Schistosoma mansoni/genetics, MESH: Pyrrolidines/chemical synthesis
Abstract: International audience; A promising means in the search of new small molecules for the treatment of schistosomiasis (amongst other parasitic ailments) is by targeting the parasitic epigenome. In the present study, a docking based virtual screening procedure using the crystal structure of histone deacetylase 8 from Schistosoma mansoni (smHDAC8) was designed. From the developed screening protocol, we were able to identify eight novel N-(2,5-dioxopyrrolidin-3-yl)-n-alkylhydroxamate derivatives as smHDAC8 inhibitors with IC50 values ranging from 4.4-20.3 µM against smHDAC8. These newly identified inhibitors were further tested against human histone deacetylases (hsHDAC1, 6 and 8), and were found also to be exerting interesting activity against them. In silico prediction of the docking pose of the compounds was confirmed by the resolved crystal structure of one of the identified hits. This confirmed these compounds were able to chelate the catalytic zinc ion in a bidentate fashion, whilst showing an inverted binding mode of the hydroxamate group when compared to the reported smHDAC8/hydroxamates crystal structures. Therefore, they can be considered as new potential scaffold for the development of new smHDAC8 inhibitors by further investigation of their structure-activity relationship.
Published: 2018

23. 8-Aryl-6-chloro-3-nitro-2-(phenylsulfonylmethyl)imidazo[1,2-a] pyridines as potent antitrypanosomatid molecules bioactivated by type 1 nitroreductases

Author: Maxime D. Crozet, Alan H. Fairlamb, Geneviève Pratviel, Alexis Valentin, Pascal Rathelot, Alix Sournia-Saquet, Clotilde Boudot, Anita Cohen, Patrice Vanelle, Julien Pedron, Jean-Luc Stigliani, Sandra Bourgeade-Delmas, Susan Wyllie, Pierre Verhaeghe, Nadine Azas, Caroline Castera-Ducros, Cyril Fersing, Alain Moreau, Sébastien Hutter, Nicolas Primas, Bertrand Courtioux, Institut de Chimie Radicalaire (ICR), Aix Marseille Université (AMU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Neuroépidémiologie Tropicale (NET), CHU Limoges-Institut d'Epidémiologie Neurologique et de Neurologie Tropicale-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST), Université de Limoges (UNILIM)-Université de Limoges (UNILIM), Laboratoire de chimie de coordination (LCC), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Infections Parasitaires : Transmission, Physiopathologie et Thérapeutiques (IP-TPT), Service de Santé des Armées-Assistance Publique - Hôpitaux de Marseille (APHM)-Aix Marseille Université (AMU)-Institut de Recherche pour le Développement (IRD), Aix-Marseille Université - Faculté de pharmacie (AMU PHARM), Aix Marseille Université (AMU), Pharmacochimie et Biologie pour le Développement (PHARMA-DEV), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie de Toulouse (ICT-FR 2599), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut de Chimie du CNRS (INC)-Institut de Recherche pour le Développement (IRD), Department of Biochemistry, Medical Sciences Institute, Dundee University, Dundee, Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Assistance Publique - Hôpitaux de Marseille (APHM)-Service de Santé des Armées, Institut de Recherche pour le Développement (IRD)-Institut de Chimie de Toulouse (ICT), and Université de Toulouse (UT)
Subjects: Models, Molecular, 0301 basic medicine, SARs, Redox potentials, HepG2 cytotoxicity, Crystallography, X-Ray, MESH: Parasitic Sensitivity Tests, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Parasitic Sensitivity Tests, In vitro activity, Drug Discovery, Nitroheterocycles, Moiety, MESH: Trypanocidal Agents, Imidazo[1, Leishmania, 2-a]pyridine, Molecular Structure, MESH: Drug Screening Assays, Antitumor, Hep G2 Cells, General Medicine, Trypanocidal Agents, 3. Good health, MESH: Nitroreductases, MESH: Cell Survival, Imidazo[1,2-a]pyridine, Pharmacophore, MESH: Models, Molecular, Trypanosoma, MESH: Cell Line, Tumor, Cell Survival, Stereochemistry, Antiparasitic, medicine.drug_class, Trypanosoma brucei brucei, 030106 microbiology, MESH: Molecular Structure, Substituent, Antineoplastic Agents, MESH: Hep G2 Cells, Article, Structure-Activity Relationship, 03 medical and health sciences, Cell Line, Tumor, MESH: Cell Proliferation, medicine, Humans, Amastigote, Cell Proliferation, Pharmacology, MESH: Humans, MESH: Leishmania donovani, Dose-Response Relationship, Drug, Aryl, Organic Chemistry, MESH: Trypanosoma brucei brucei, Nitroreductases, MESH: Crystallography, X-Ray, 030104 developmental biology, chemistry, Nitro, Suzuki-Miyaura cross-coupling reaction, MESH: Antineoplastic Agents, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Drug Screening Assays, Antitumor, Leishmania donovani, Fexinidazole
Abstract: International audience; Based on a previously identified antileishmanial 6,8-dibromo-3-nitroimidazo[1,2-a]pyridine derivative, a Suzuki-Miyaura coupling reaction at position 8 of the scaffold was studied and optimized from a 8-bromo-6-chloro-3-nitroimidazo[1,2-a]pyridine substrate. Twenty-one original derivatives were prepared, screened in vitro for activity against L. infantum axenic amastigotes and T. brucei brucei trypomastigotes and evaluated for their cytotoxicity on the HepG2 human cell line. Thus, 7 antileishmanial hit compounds were identified, displaying IC50 values in the 1.1-3 μM range. Compounds 13 and 23, the 2 most selective molecules (SI = >18 or >17) were additionally tested on both the promastigote and intramacrophage amastigote stages of L. donovani. The two molecules presented a good activity (IC50 = 1.2-1.3 μM) on the promastigote stage but only molecule 23, bearing a 4-pyridinyl substituent at position 8, was active on the intracellular amastigote stage, with a good IC50 value (2.3 μM), slightly lower than the one of miltefosine (IC50 = 4.3 μM). The antiparasitic screening also revealed 8 antitrypanosomal hit compounds, including 14 and 20, 2 very active (IC50 = 0.04-0.16 μM) and selective (SI = >313 to 550) molecules toward T. brucei brucei, in comparison with drug-candidate fexinidazole (IC50 = 0.6 & SI > 333) or reference drugs suramin and eflornithine (respective IC50 = 0.03 and 13.3 μM). Introducing an aryl moiety at position 8 of the scaffold quite significantly increased the antitrypanosomal activity of the pharmacophore. Antikinetoplastid molecules 13, 14, 20 and 23 were assessed for bioactivation by parasitic nitroreductases (either in L. donovani or in T. brucei brucei), using genetically modified parasite strains that over-express NTRs: all these molecules are substrates of type 1 nitroreductases (NTR1), such as those that are responsible for the bioactivation of fexinidazole. Reduction potentials measured for these 4 hit compounds were higher than that of fexinidazole (-0.83 V), ranging from -0.70 to -0.64 V.
Published: 2018

24. Functional mapping of factor VIII C2 domain

Author: Marc Delcourt, Jean-Luc Pellequer, Jean-Luc Plantier, Didier Saboulard, Claude Negrier, Shu-wen W. Chen, Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Service de Biochimie et Toxicologie Nucléaire (SBTN), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), BIOMETHODES, Centre Hospitalier Universitaire de Lyon (CHU Lyon), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Mutant, MESH: Protein Structure, Secondary, 030204 cardiovascular system & hematology, Epitope, Protein Structure, Secondary, law.invention, MESH: Protein Structure, Tertiary, 0302 clinical medicine, MESH: Structure-Activity Relationship, law, MESH: Chlorocebus aethiops, hemic and lymphatic diseases, Chlorocebus aethiops, MESH: Animals, MESH: von Willebrand Factor, C2 domain, Alanine, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Chemistry, Hematology, MESH: Factor VIII, MESH: COS Cells, MESH: Mutagenesis, Site-Directed, Biochemistry, MESH: Blood Coagulation, COS Cells, Recombinant DNA, Blood Coagulation Tests, Protein Binding, MESH: Mutation, MESH: Alanine, In silico, Cofactor, 03 medical and health sciences, Structure-Activity Relationship, Von Willebrand factor, von Willebrand Factor, Animals, Humans, MESH: Protein Binding, Blood Coagulation, Factor VIII, MESH: Humans, MESH: Blood Coagulation Tests, Protein Structure, Tertiary, 030104 developmental biology, Mutation, biology.protein, Mutagenesis, Site-Directed
Abstract: SummaryThe factor VIII (FVIII) is a cofactor of the coagulation cascade. The FVIII C2 domain is a critical domain that participates in the interactions with the von Willebrand factor and the phospholipidic surfaces. To assess the importance of each residue of this domain in the maintenance of the structure and the function of FVIII, a number (n=139) of mutants were generated by substituting the original residues, from Ser2173 to Gly2325, by an alanine. Mutants were built within a complete B domain- deleted FVIII and expressed in COS-1 cells. Mutant antigen levels and procoagulant activities were measured. Two in silico analyses, a sliding average procedure and an analysis of the mutation energy cost were conducted in parallel on the FVIII structure. Both results were in agreement with the functional data, and illustrated the benefit of using such strategies prior to targeting specific residues in the aim of generating active recombinant molecules. The functional assays identify the residues that are important to maintaining the structure of the C2 domain, mainly those forming β-sheet, and those that can afford substitution, establishing a detailed functional relation with the available crystallographic data. This study provided a comprehensive functional mapping of the FVIII C2 domain and discussed the implication of specific residues in respect to the maintenance in the activity and structure stability, the efficiency in secretion, the binding to phospholipids and the formation of epitope.
Published: 2017

25. Evidence of K+ homeostasis disruption in cellular dysfunction triggered by 7-ketocholesterol, 24S-hydroxycholesterol, and tetracosanoic acid (C24:0) in 158N murine oligodendrocytes

Author: Jérôme De Seze, Thomas Nury, Gérard Lizard, Mustapha Cherkaoui-Malki, Quentin Raas, Mohamed El-Ayeb, Meryam Debbabi, Mohammad Samadi, Anne Vejux, Maryem Bezine, Rym Ben-Khalifa, Thibault Moreau, Laboratoire Bio-PeroxIL. Biochimie du peroxysome, inflammation et métabolisme lipidique [Dijon] (BIO-PEROXIL), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Université de Monastir - University of Monastir (UM), Laboratoire des Venins et Biomolécules Thérapeutiques - Laboratory of Venoms and Therapeutic Biomolecules (LR11IPT08), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Laboratoire de Chimie et Physique - Approche Multi-échelle des Milieux Complexes (LCP-A2MC), Université de Lorraine (UL), Hôpital de Hautepierre [Strasbourg], Service de Neurologie générale, vasculaire et dégénérative (CHU de Dijon), and Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)
Subjects: 0301 basic medicine, MESH: Mice, Mutant Strains, [SDV]Life Sciences [q-bio], Cell, Biochemistry, MESH: Dose-Response Relationship, Drug, 03 medical and health sciences, Tetracosanoic acid (C24:0), MESH: Hydroxycholesterols, 0302 clinical medicine, MESH: Structure-Activity Relationship, MESH: Cell Proliferation, medicine, MESH: Animals, Potassium kv channels, 4-Aminopyridine, Lipid bilayer, Cell adhesion, Molecular Biology, MESH: Mice, Membrane potential, Cell growth, Chemistry, Organic Chemistry, 158N murine oligodendrocytes, MESH: Ketocholesterols, Depolarization, MESH: Oligodendroglia, Cell Biology, 7-Ketocholesterol, MESH: Male, 3. Good health, Cell biology, MESH: Fatty Acids, 030104 developmental biology, medicine.anatomical_structure, MESH: Homeostasis, 24S-Hydroxycholesterol, MESH: Potassium, sense organs, 030217 neurology & neurosurgery, Intracellular, Homeostasis, MESH: Cells, Cultured
Abstract: International audience; Imbalance in the homeostasis of K+ ions has been reported to contribute to the pathogenesis of neurodegenerative diseases. 7-ketocholesterol (7KC), 24S-hydroxycholesterol (24S-OHC), and tetracosanoic acid (C24:0), often found at increased levels in patients with Alzheimer's disease, Multiple Sclerosis and X-ALD, are able to trigger numerous nerve cell dysfunctions. We therefore studied the impact of 7KC, 24S-OHC, and C24:0 on 158N murine oligodendrocytes, and determined their impact on K+ homeostasis. The effects of 7KC, 24S-OHC and C24:0 on lipid membrane organization and membrane potential were examined with merocyanine 540 (MC540) and bis-(1,3-diethylthiobarbituric acid) trimethine oxonol (DiSBAC2(3)), respectively. The intracellular concentration of K+ ([K+]i) was measured by flame photometry and the ratiometric approach using the PBFI-AM fluorescence indicator. To determine the relationships between [K+]i and lipotoxicity, 158N cells were pre-treated with a universal Kv channels blocker, 4-aminopyridine (4-AP), without or with 7KC, 24S-OHC or C24:0. Cell adhesion, cell growth, mitochondrial depolarization, cytoplasmic membrane integrity, the presence of SubG1 and the morphological aspect of the nuclei were determined with various microscopy, flow cytometry and biochemistry methods. 7KC, 24S-OHC and C24:0 induced changes in lipid content and polarization of the cytoplasmic membrane. These events were associated with increased [K+]i. Blocking Kv channels with 4-AP exacerbated 7KC-, 24S-OHC- and C24:0-induced cell dysfunction. 4-AP exacerbated loss of cell adhesion and cell growth inhibition, amplified mitochondrial depolarization and cytoplasmic membrane damage, and increased the percentage of SubG1 cells. The positive correlation between [K+]i and cell death supports the potential involvement of K+ in 7KC-, 24S-OHC-, and C24:0-induced cytotoxicity.
Published: 2017

26. The spread of prion-like proteins by lysosomes and tunneling nanotubes: Implications for neurodegenerative diseases

Author: Chiara Zurzolo, Guiliana Soraya Victoria, Trafic membranaire et Pathogénèse, Institut Pasteur [Paris], C. Zurzolo was supported by the Agence Nationale de Recherche (grants Joint Programme–Neurodegenerative Disease Neutargets: ANR-14-JPCD-0002-01 and ANR-16 CE160019-01 NEUROTUNN) and the Equipe Fondation Recherche Médicale 2014 (grant DEQ20140329557). G. Soraya Victoria was supported by Bourse Pasteur-Roux, Institut Pasteur, Paris., We thank Drs. Ayşegül Dilsizoğlu-Şenol, Yuan-Ju Wu, and Frida Loria Salinas for their critical reading of the manuscript., ANR-16-CE16-0019,Neurotunn,Role des nanotubes membranaires dans la propagation d'agrégats protéiques impliqués dans les maladie neurodégénératives(2016), ANR-14-JPCD-0002,Neutargets,Targeting the propagation of pathogenic protein assemblies in neurodegenerative disease(2014), Victoria, G. S., Zurzolo, C., and Institut Pasteur [Paris] (IP)
Subjects: 0301 basic medicine, MESH: Signal Transduction, Protein Folding, Protein Conformation, animal diseases, Cell, Review, Protein aggregation, MESH: Nerve Degeneration, Proteostasis Deficiencie, MESH: Neurodegenerative Diseases, Protein structure, MESH: Protein Conformation, MESH: Structure-Activity Relationship, MESH: Animals, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH: Proteostasis Deficiencies, Nanotubes, MESH: Protein Aggregation, Pathological, Neurodegenerative Diseases, GPI-Linked Protein, Lysosome, 3. Good health, Cell biology, Transport protein, Protein Transport, medicine.anatomical_structure, Prion, Protein folding, Signal transduction, MESH: Nanotubes, Intracellular, Human, Signal Transduction, MESH: Protein Transport, Prions, MESH: Protein Folding, Reviews, Biology, GPI-Linked Proteins, Protein Aggregation, Pathological, 03 medical and health sciences, Protein Aggregates, Structure-Activity Relationship, MESH: Prions, medicine, Animals, Humans, Prion protein, Proteostasis Deficiencies, MESH: Humans, Neurodegenerative Disease, Animal, Cell Biology, nervous system diseases, MESH: Protein Aggregates, 030104 developmental biology, Nerve Degeneration, Protein Aggregate, MESH: GPI-Linked Proteins, Lysosomes, MESH: Lysosomes
Abstract: Victoria and Zurzolo discuss current evidence for the emerging role of lysosomal damage and tunneling nanotubes in the intercellular propagation of prion and prion-like proteins in neurodegenerative disease., Progression of pathology in neurodegenerative diseases is hypothesized to be a non–cell-autonomous process that may be mediated by the productive spreading of prion-like protein aggregates from a “donor cell” that is the source of misfolded aggregates to an “acceptor cell” in which misfolding is propagated by conversion of the normal protein. Although the proteins involved in the various diseases are unrelated, common pathways appear to be used for their intercellular propagation and spreading. Here, we summarize recent evidence of the molecular mechanisms relevant for the intercellular trafficking of protein aggregates involved in prion, Alzheimer’s, Huntington’s, and Parkinson’s diseases. We focus in particular on the common roles that lysosomes and tunneling nanotubes play in the formation and spreading of prion-like assemblies.
Published: 2017

27. BF 2 -azadipyrromethene NIR-emissive fluorophores with research and clinical potential

Author: Gonzalo Sampedro, Ghazi Ismail, Harrison C. Daly, Donal F. O'Shea, Ronan Cahill, Corentin Bon, Dan Wu, Royal College of Surgeons in Ireland (RCSI), and University College Dublin [Dublin] (UCD)
Subjects: Azadipyrromethene, Fluorescence-lifetime imaging microscopy, Fluorescence guided surgery, Fluorophore, MESH: Porphobilinogen, [SDV]Life Sciences [q-bio], MESH: Boron Compounds, MESH: Molecular Structure, Analytical chemistry, 010402 general chemistry, 01 natural sciences, Tattoo, MESH: Spectroscopy, Near-Infrared, MESH: Dose-Response Relationship, Drug, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, MESH: Structure-Activity Relationship, Drug Discovery, Bathochromic shift, [CHIM]Chemical Sciences, Clinical imaging, MESH: Colon, Pharmacology, MESH: Humans, Organic Chemistry, Near-infrared spectroscopy, General Medicine, NIR-fluorescence, MESH: Fluorescent Dyes, Fluorescence, 0104 chemical sciences, chemistry, 030220 oncology & carcinogenesis, MESH: Aza Compounds, Lymph node, Preclinical imaging, Biomedical engineering
Abstract: International audience; The use of near-infrared fluorescence for in vivo research and intraoperative clinical imaging is rapidly expanding, with new applications being proposed and developed. While imaging hardware and software have significantly progressed in recent times, the molecular fluorescent agents remain a limiting factor. In this report, the design, synthesis, photophysical characterization and bio-medical imaging assessment of two new NIR-fluorophores based on the BF2-azadipyrromethene fluorophore class are described. Inclusion of dimethylamino substituents on these BF2-azadipyrromethene probes results in very large bathochromic shifts with photophysical measurements showing absorption and emission maxima between 757 and 818 nm within the desired NIR spectra region. Testing of the probes shows that they are suitable for fluorescence imaging with both research and clinical instrumentation. Preclinical imaging assessment shows their suitability as fluorescent markers (tattoos) of lesions for intraoperative identification and lymphatic mapping in ex vivo human colonic tissue. These new clinical wavelength-compatible fluorophores may contribute towards the on-going expansion of medical uses for NIR-fluorescence.
Published: 2017

28. Evolved distal tail carbohydrate binding modules of L actobacillus phage J-1: a novel type of anti-receptor widespread among lactic acid bacteria phages

Author: Dieterle, Maria-Eugenia, Spinelli, Silvia, Sadovskaya, Irina, Piuri, Mariana, Cambillau, Christian, Architecture et fonction des macromolécules biologiques (AFMB), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Instituto de Química Biológica Ciencias Exactas y Naturales (IQUIBICEN), Consejo Nacional de Investigaciones Científicas y Técnicas [Buenos Aires] (CONICET)-Departamento de Quimica Biológica, Facultad de Ciencias Exactas y Naturales [Buenos Aires] (FCEyN), Universidad de Buenos Aires [Buenos Aires] (UBA)-Universidad de Buenos Aires [Buenos Aires] (UBA)-Facultad de Ciencias Exactas y Naturales [Buenos Aires] (FCEyN), Universidad de Buenos Aires [Buenos Aires] (UBA)-Universidad de Buenos Aires [Buenos Aires] (UBA), LR2B UMT 08, Bassin Napoleon, Univ Littoral Cote dOpale, Université du Littoral Côte d'Opale (ULCO), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA), Departamento de Quimica Biológica, and Universidad de Buenos Aires [Buenos Aires] (UBA)-Universidad de Buenos Aires [Buenos Aires] (UBA)-Consejo Nacional de Investigaciones Científicas y Técnicas [Buenos Aires] (CONICET)
Subjects: [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH: Host Specificity, Protein Conformation, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Carbohydrates, MESH: Microscopy, Electron, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Host Specificity, Structure-Activity Relationship, MESH: Protein Conformation, MESH: Structure-Activity Relationship, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Protein Binding, Bacteriophages, Lactic Acid, MESH: Bacteriophages, MESH: Viral Tail Proteins, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MESH: Lactobacillus casei, Virion, Viral Tail Proteins, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Lactococcus lactis, Lactobacillus, Lacticaseibacillus casei, Microscopy, Electron, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], MESH: Lactococcus lactis, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, MESH: Virion, MESH: Lactic Acid, MESH: Lactobacillus, Protein Binding, MESH: Carbohydrates
Abstract: International audience; Bacteriophage replication requires specific host-recognition. Some siphophages harbour a large complex, the baseplate, at the tip of their non-contractile tail. This baseplate holds receptor binding proteins (RBPs) that can recognize the host cell-wall polysaccharide (CWPS) and specifically attach the phage to its host. While most phages possess a dedicated RBP, the phage J-1 that infects Lactobacillus casei seemed to lack one. It has been shown that the phage J-1 distal tail protein (Dit) plays a role in host recognition and that its sequence comprises two inserted modules compared with 'classical' Dits. The first insertion is similar to carbohydrate-binding modules (CBMs), whereas the second insertion remains undocumented. Here, we determined the structure of the second insertion and found it also similar to several CBMs. Expressed insertion CBM2, but not CBM1, binds to L. casei cells and neutralize phage attachment to the bacterial cell wall and the isolated and purified CWPS of L. casei BL23 prevents CBM2 attachment to the host. Electron microscopy single particle reconstruction of the J-1 virion baseplate revealed that CBM2 is projected at the periphery of Dit to optimally bind the CWPS receptor. Taken together, these results identify J-1 evolved Dit as the phage RBP.
Published: 2017

29. Identification of cancer-associated missense mutations in hace1 that impair cell growth control and Rac1 ubiquitylation

Author: Emilie Andrio, Emmanuel Lemichez, Jacqueline Cherfils, Poul H. Sorensen, Anne Doye, Daniel Hamaoui, Mads Daugaard, Raymond Ruimy, Frédéric Bost, Amel Mettouchi, Romain Lotte, Centre méditerranéen de médecine moléculaire (C3M), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Toxines bactériennes dans la relation hôtes-pathogènes, Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR50-Université Côte d'Azur (UCA)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), Laboratoire de Bactériologie [CHU Nice], Centre Hospitalier Universitaire de Nice (CHU Nice)-Hôpital Archet 2 [Nice] (CHU), Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), University of British Columbia (UBC), Vancouver Prostate Centre [Vancouver General Hospital] (VPC), Vancouver General Hospital, BC Cancer Agency Research Centre (BCCRC), Signalisation moléculaire et obésité, COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-IFR50-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by INSERM, CNRS and fundings from the Ligue Nationale Contre le Cancer (LNCC, équipe labellisée), Association pour la Recherche sur le Cancer (ARC-SFI20111203659 & ARC-SFI20111203671), a fellowship LNCC to D.H., a PhD fellowship from the 'Fondation pour la Recherche Médicale' (FRM-FDM20150632804) to R.L., We are grateful to Melissa Sivaneson, Frédéric Reinier and Patrick Munro for technical help and fruitful discussions. We thank Orane Visvikis for critical reading of the manuscript., Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Côte d'Azur (UCA), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-IFR50-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Côte d'Azur (UCA)
Subjects: Models, Molecular, rac1 GTP-Binding Protein, 0301 basic medicine, HECT domain, MESH: Ubiquitin-Protein Ligases/chemistry, Somatic cell, [SDV]Life Sciences [q-bio], MESH: Amino Acid Sequence, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], MESH: Structure-Activity Relationship, Ubiquitin, Neoplasms, MESH: Ubiquitin-Protein Ligases/genetics, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], MESH: Ubiquitin-Protein Ligases/metabolism, Ankyrin, Small GTPase, Genetics, chemistry.chemical_classification, Multidisciplinary, biology, 3. Good health, Ubiquitin ligase, Cell biology, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, MESH: Protein Domains, MESH: Models, Molecular, Protein Binding, MESH: rac1 GTP-Binding Protein/metabolism, MESH: Mutant Proteins/chemistry, Ubiquitin-Protein Ligases, Protein domain, Mutation, Missense, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Article, Cell Line, MESH: Mutation, Missense/genetics, Structure-Activity Relationship, 03 medical and health sciences, Protein Domains, MESH: Neoplasms/genetics, MESH: Cell Proliferation, Humans, MESH: Protein Binding, Amino Acid Sequence, Cell Proliferation, MESH: Humans, Cell growth, Ubiquitination, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: Cell Line, 030104 developmental biology, chemistry, Proteolysis, biology.protein, Mutant Proteins, MESH: Ubiquitination
Abstract: The E3 ubiquitin ligase HACE1 is a potent tumor suppressor that controls cell proliferation and ubiquitylates the small GTPase Rac1 to target it to proteasomal degradation. Whether and how the activity of HACE1 is regulated by the N-terminal ankyrin (ANK) and the middle (MID) domains is ill defined. Here, we identified in the version 64 of the Catalogue of Somatic Mutations in Cancer (COSMIC) 13 missense mutations of hace1 located outside the HECT domain, and found that all lead to defective control of cell proliferation. In addition, several mutations located in the ankyrin domain displayed a dramatic reduction in Rac1 ubiquitylation associated with a decrease of colony formation in soft agar. 3D structure modelling of the 7 ankyrin-repeats coupled to functional analysis identified a surface epitope centered on one of the mutated residue, Gly-175, which is critical for controlling Rac1 binding and ubiquitylation. We also identified a role for the MID domain in conferring the specificity of association of HACE1 to the active form of Rac1. Our study of the functional interplay between HACE1 and Rac1 in cancer thus sheds a new light on the molecular mechanism of Rac1 ubiquitylation by HACE1 and the impact of its cancer-associated mutations in cell proliferation.
Published: 2017

30. Introns Protect Eukaryotic Genomes from Transcription-Associated Genetic Instability

Author: Vincent Géli, Ana Rita Grosso, Guilhem Janbon, Emeline Coleno, Sérgio F. de Almeida, Amandine Bonnet, Adrien Presle, Sree Rama Chaitanya Sridhara, Abdessamad El-Kaoutari, Benoit Palancade, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Faculdade de Medicina [Lisboa], Universidade de Lisboa = University of Lisbon (ULISBOA), Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Biologie des ARN des Pathogènes fongiques - RNA Biology of Fungal Pathogens, Institut Pasteur [Paris] (IP), This work was supported by CNRS (to B.P.), Fondation ARC pour la Recherche sur le Cancer (to B.P.), Ligue Nationale contre le Cancer (to B.P., fellowship to A.B. and Equipe Labellisée 2014 to V.G.), EMBO (short-term fellowship to A.B.), Cancéropole PACA (fellowship to A.E.), and Fundaçao para a Ciencia e Tecnologia, Portugal (PTDC/BIM-ONC/0016-2014 to S.F.d.A. and IF/00510/2014 to A.R.G.). Bioinformatic and computing support at CRCM was provided by the CRCM Integrative Bioinformatics and Datacentre IT and Scientific Computing platforms., Universidade de Lisboa (ULISBOA), Institut Pasteur [Paris], Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU), and Biologie des ARN des Pathogènes fongiques
Subjects: 0301 basic medicine, Transcription, Genetic, MESH: Introns, [SDV]Life Sciences [q-bio], Candida glabrata, mRNA splicing, MESH: Genotype, 0302 clinical medicine, MESH: Structure-Activity Relationship, Minor spliceosome, Gene Expression Regulation, Fungal, Databases, Genetic, DNA, Fungal, MESH: Candida glabrata, MESH: Databases, Genetic, Genetics, Splice site mutation, MESH: Genomic Instability, Nucleic Acid Heteroduplexes, genetic instability, MESH: Cryptococcus neoformans, Group II intron, R-loops, MESH: Saccharomyces cerevisiae, messenger ribonucleoparticle, Phenotype, MESH: Nucleic Acid Conformation, MESH: Schizosaccharomyces, Ribonucleoproteins, RNA splicing, MESH: Fungal Proteins, transcription, MESH: Spliceosomes, MESH: Gene Expression Regulation, Fungal, MESH: Computational Biology, Spliceosome, Genotype, intron, RNA Splicing, Saccharomyces cerevisiae, Biology, MESH: Phenotype, Genomic Instability, Cell Line, Fungal Proteins, Structure-Activity Relationship, 03 medical and health sciences, mRNP, MESH: Nucleic Acid Heteroduplexes, Schizosaccharomyces, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecular Biology, Gene, MESH: DNA Damage, MESH: Humans, MESH: RNA, Fungal, MESH: Transcription, Genetic, Intron, Computational Biology, RNA, RNA, Fungal, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Biology, Introns, recombination, MESH: Cell Line, MESH: DNA, Fungal, MESH: Ribonucleoproteins, 030104 developmental biology, Cryptococcus neoformans, Spliceosomes, Nucleic Acid Conformation, MESH: RNA Splicing, 030217 neurology & neurosurgery, DNA Damage
Abstract: International audience; Transcription is a source of genetic instability that can notably result from the formation of genotoxic DNA:RNA hybrids, or R-loops, between the nascent mRNA and its template. Here we report an unexpected function for introns in counteracting R-loop accumulation in eukaryotic genomes. Deletion of endogenous introns increases R-loop formation, while insertion of an intron into an intronless gene suppresses R-loop accumulation and its deleterious impact on transcription and recombination in yeast. Recruitment of the spliceosome onto the mRNA, but not splicing per se, is shown to be critical to attenuate R-loop formation and transcription-associated genetic instability. Genome-wide analyses in a number of distant species differing in their intron content, including human, further revealed that intron-containing genes and the intron-richest genomes are best protected against R-loop accumulation and subsequent genetic instability. Our results thereby provide a possible rationale for the conservation of introns throughout the eukaryotic lineage.
Published: 2017
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31. 9- and 11-substituted 4-azapaullones are potent and selective inhibitors of African trypanosoma

Author: Mahin Abad Dar, Franziska Maiwald, Diego Benítez, Laurent Meijer, Nadège Loaëc, Conrad Kunick, Lutz Preu, Hanna Erdmann, Marcelo A. Comini, Oliver Koch, Diego Charquero, Christoph Hölscher, Technische Universität Braunschweig = Technical University of Braunschweig [Braunschweig], Institut Pasteur de Montevideo, Réseau International des Instituts Pasteur (RIIP), Forschungszentrum Borstel - Research Center Borstel, Technische Universität Dortmund [Dortmund] (TU), ManRos Therapeutics, This project was funded by the German 'Bundesministerium für Bildung und Forschung' (KMU-innovativ 5, Förderkennzeichen 0315814, to F. M., M. A. D., O. K., C. H., and C. K.). The support of FOCEM (MERCOSUR Structural Convergence Fund, COF 03/11) and ANII (grant Innova Uruguay, agreement DCI-ALA/2007/19.040 between Uruguay and the European Commission, and INI_X_2011_1_4077 and POS_NAC_2013_1_114477) is acknowledged by M. A. C., D. B. and D. C.. L. M. was supported by a grant from the ‘Agence Nationale de la Recherche’ (ANR- 11-RPIB-0016 grant, Transleish).
Subjects: MESH: Trypanocidal Agents/chemical synthesis, MESH: Benzazepines/toxicity, Nagana, Mice, Chalcones, MESH: Structure-Activity Relationship, MESH: Cell Proliferation/drug effects, Human trypanosomiasis, Drug Discovery, MESH: Animals, MESH: Benzazepines/chemistry, biology, MESH: Trypanosoma brucei brucei/physiology, Chemistry, MESH: Macrophages/drug effects, General Medicine, Trypanocidal Agents, 3. Good health, MESH: Trypanocidal Agents/pharmacology, Biochemistry, Paullones, MESH: Trypanocidal Agents/toxicity, MESH: Trypanosomiasis, African/parasitology, Trypanosoma brucei brucei, Cell Line, Structure-Activity Relationship, Heck reaction, parasitic diseases, medicine, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Trypanosoma brucei brucei/drug effects, MESH: Drug Design, MESH: Mice, Cell Proliferation, Pharmacology, MESH: Humans, MESH: Benzazepines/chemical synthesis, MESH: Trypanocidal Agents/chemistry, Macrophages, Organic Chemistry, MESH: Benzazepines/pharmacology, Benzazepines, biology.organism_classification, medicine.disease, Antiparasitic agent, Cinnamides, MESH: Cell Line, Trypanosomiasis, African, Drug Design, Trypanosoma
Abstract: International audience; Trypanosomes from the "brucei" complex are pathogenic parasites endemic in sub-Saharan Africa and causative agents of severe diseases in humans and livestock. In order to identify new antitrypanosomal chemotypes against African trypanosomes, 4-azapaullones carrying α,β-unsaturated carbonyl chains in 9- or 11-position were synthesized employing a procedure with a Heck reaction as key step. Among the so prepared compounds, 5a and 5e proved to be potent antiparasitic agents with antitrypanosomal activity in the submicromolar range.
Published: 2014

32. Evaluation of the antiprion activity of 6-aminophenanthridines and related heterocycles

Author: Justine Evrard, Flavie Soubigou, Cécile Voisset, Nathalie Desban, Pascale Lemoine, Nassima Oumata, Marc Blondel, Serge Bouaziz, Phuhai Nguyen, Génétique, génomique fonctionnelle et biotechnologies (UMR 1078) (GGB), Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Université de Brest (UBO)-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), ManRos Therapeutics, Systèmes d'Elevage, Nutrition Animale et Humaine (SENAH), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Supérieure Agronomique de Rennes, Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), EFS-Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), INSERM Unit 1018, Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC), Bouaziz, Serge, Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Models, Molecular, MESH: Heterocyclic Compounds, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Prions, MESH: Mice, Transgenic, MESH: Molecular Structure, Cell, Mice, Transgenic, Saccharomyces cerevisiae, Mice, Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, MESH: Prions, MESH: Structure-Activity Relationship, 0302 clinical medicine, Heterocyclic Compounds, Mammalian cell, Drug Discovery, medicine, Animals, MESH: Animals, MESH: Mice, IC50, Phenanthridine, 030304 developmental biology, Pharmacology, 0303 health sciences, Molecular Structure, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Neurodegenerative diseases, Organic Chemistry, Aromaticity, General Medicine, MESH: Phenanthridines, MESH: Saccharomyces cerevisiae, Yeast, Phenanthridines, 3. Good health, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], medicine.anatomical_structure, chemistry, Biochemistry, Protein folding, MESH: Models, Molecular, 030217 neurology & neurosurgery, Ex vivo, Protein misfolding
Abstract: International audience; Series of 6-aminophenanthridines and related heterocyclic compounds such as benzonaphtyridines were prepared. Reduction of one of the three aromatic rings was also performed. The compounds were first tested for their antiprion activity in a previously described yeast-based colourimetric prion assay. The most potent derivatives were then assayed ex vivo against the mammalian prion PrP(Sc) in a cell-based assay. Several of the new compounds were found more potent than the parent lead 6-aminophenanthridine. The most promising compounds against yeast and mammalian prions were 8-azido-6-aminophenanthridine (3m), and 7,10-dihydrophenanthridin-6-amine (14). In the mammalian cell-based assay, the IC50 of these two compounds were around 5 μM and 1.8 μM, respectively.
Published: 2014

33. MOLECULAR EVOLUTION OF GPCRS: Kisspeptin/kisspeptin receptors

Author: Jérémy Pasquier, Nédia Kamech, Karine Rousseau, Anne-Gaëlle Lafont, Hubert Vaudry, Sylvie Dufour, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), and Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)
Subjects: MESH: Signal Transduction, receptor, MESH: Amino Acid Sequence, MESH: Receptors, G-Protein-Coupled, Bioinformatics, Receptors, G-Protein-Coupled, MESH: Structure-Activity Relationship, 0302 clinical medicine, Endocrinology, Kisspeptin, Gene Duplication, Gene duplication, MESH: Animals, MESH: Genetic Variation, Neoplasm Metastasis, MESH: Evolution, Molecular, Conserved Sequence, Kisspeptins, 0303 health sciences, MESH: Conserved Sequence, MESH: Kisspeptins, biology, Phylogenetic tree, MESH: Gene Duplication, Vertebrate, MESH: Receptors, Kisspeptin-1, Biological Evolution, Signal Transduction, endocrine system, MESH: Biological Evolution, kisspeptin, Evolution, Molecular, Structure-Activity Relationship, 03 medical and health sciences, Molecular evolution, biology.animal, evolution, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Molecular Biology, Gene, 030304 developmental biology, MESH: Humans, Genetic Variation, MESH: Neoplasm Metastasis, Evolutionary biology, vertebrates, human activities, 030217 neurology & neurosurgery, Function (biology), Receptors, Kisspeptin-1
Abstract: Following the discovery of kisspeptin (Kiss) and its receptor (GPR54 or KissR) in mammals, phylogenetic studies revealed up to three Kiss and four KissR paralogous genes in other vertebrates. The multiplicity of Kiss and KissR types in vertebrates probably originated from the two rounds of whole-genome duplication (1R and 2R) that occurred in early vertebrates. This review examines compelling recent advances on molecular diversity and phylogenetic evolution of vertebrate Kiss and KissR. It also addresses, from an evolutionary point of view, the issues of the structure–activity relationships and interaction of Kiss with KissR and of their signaling pathways. Independent gene losses, during vertebrate evolution, have shaped the repertoire ofKissandKissRin the extant vertebrate species. In particular, there is no conserved combination of a givenKisstype with aKissRtype, across vertebrate evolution. The striking conservation of the biologically active ten-amino-acid C-terminal sequence of all vertebrate kisspeptins, probably allowed this evolutionary flexibility of Kiss/KissR pairs. KissR mutations, responsible for hypogonadotropic hypogonadism in humans, mostly occurred at highly conserved amino acid positions among vertebrate KissR. This further highlights the key role of these amino acids in KissR function. In contrast, less conserved KissR regions, notably in the intracellular C-terminal domain, may account for differential intracellular signaling pathways between vertebrate KissR. Cross talk between evolutionary and biomedical studies should contribute to further understanding of the Kiss/KissR structure–activity relationships and biological functions.
Published: 2014

34. Structure–activity relationships of new cyanothiophene inhibitors of the essential peptidoglycan biosynthesis enzyme MurF

Author: Stanislav Gobec, Samo Turk, Hélène Barreteau, Dominique Mengin-Lecreulx, Andréa Dessen, Alex J. O'Neill, Didier Blanot, Christopher P. Randall, Izidor Sosič, Carlos Contreras-Martel, Martina Hrast, Damijan Knez, Faculty of Pharmacy, University of Ljubljana, Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Models, Molecular, MESH: Catalytic Domain, Microbial Sensitivity Tests, Peptidoglycan, Thiophenes, MESH: Drug Design, medicine.disease_cause, 01 natural sciences, Bacterial cell structure, Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Biosynthesis, Catalytic Domain, MESH: Anti-Bacterial Agents, Drug Discovery, medicine, Nucleotide, Enzyme Inhibitors, Peptide Synthases, Escherichia coli, 030304 developmental biology, Pharmacology, chemistry.chemical_classification, MESH: Microbial Sensitivity Tests, 0303 health sciences, DNA ligase, Bacteria, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MESH: Peptidoglycan, 010405 organic chemistry, Organic Chemistry, General Medicine, MESH: Thiophenes, MESH: Peptide Synthases, Anti-Bacterial Agents, 0104 chemical sciences, MESH: Bacteria, Enzyme, chemistry, Biochemistry, MESH: Enzyme Inhibitors, Drug Design, Antibacterial activity, MESH: Models, Molecular
Abstract: International audience; Peptidoglycan is an essential component of the bacterial cell wall, and enzymes involved in its biosynthesis represent validated targets for antibacterial drug discovery. MurF catalyzes the final intracellular peptidoglycan biosynthesis step: the addition of D-Ala-D-Ala to the nucleotide precursor UDP-MurNAc-L-Ala-γ-D-Glu-meso-DAP (or L-Lys). As MurF has no human counterpart, it represents an attractive target for the development of new antibacterial drugs. Using recently published cyanothiophene inhibitors of MurF from Streptococcus pneumoniae as a starting point, we designed and synthesized a series of structurally related derivatives and investigated their inhibition of MurF enzymes from different bacterial species. Systematic structural modifications of the parent compounds resulted in a series of nanomolar inhibitors of MurF from S. pneumoniae and micromolar inhibitors of MurF from Escherichia coli and Staphylococcus aureus. Some of the inhibitors also show antibacterial activity against S. pneumoniae R6. These findings, together with two new co-crystal structures, represent an excellent starting point for further optimization toward effective novel antibacterials.
Published: 2013

35. Elaboration of a proprietary thymidylate kinase inhibitor motif towards anti-tuberculosis agents

Author: Song, Lijun, Risseeuw, Martijn D.P., Froeyen, Matheus, Karalic, Izet, Goeman, Jan, Cappoen, Davie, Van der Eycken, Johan, Cos, Paul, Munier-Lehmann, Hélène, Van Calenbergh, Serge, Risseeuw, Martijn, Froeyen, Mathy, Universiteit Gent = Ghent University [Belgium] (UGENT), Rega Institute for Medical Research [Leuven, België], Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), University of Antwerp (UA), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], This project is financially supported by China Scholarship Council (CSC)., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Universiteit Gent = Ghent University (UGENT), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Models, Molecular, MESH: Mycobacterium tuberculosis, [SDV]Life Sciences [q-bio], Clinical Biochemistry, POTENT INHIBITORS, Antitubercular Agents, Pharmaceutical Science, 01 natural sciences, Biochemistry, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, Thymidine monophosphate kinase, MESH: Structure-Activity Relationship, DESIGN, Piperidines, Drug Discovery, Medicine and Health Sciences, thymine derivatives, MESH: Protein Kinase Inhibitors, Thymidine monophosphate, MESH: Microbial Sensitivity Tests, Nucleoside-phosphate kinase, biology, Molecular Structure, Kinase, Pharmacology. Therapy, 3. Good health, Chemistry, MESH: Piperidines, ANTIBACTERIAL INHIBITORS, NUCLEOSIDE ANALOGS, Molecular Medicine, MYCOBACTERIUM-TUBERCULOSIS, MESH: Models, Molecular, medicine.drug, MESH: Molecular Structure, Microbial Sensitivity Tests, Thymidylate kinase, Mycobacterium tuberculosis, Structure-Activity Relationship, medicine, Structure–activity relationship, Humans, extensively drug-resistant tuberculosis, [CHIM]Chemical Sciences, XDR-TB, Biology, Molecular Biology, Protein Kinase Inhibitors, MESH: Humans, Nucleoside analogue, Dose-Response Relationship, Drug, 010405 organic chemistry, Inhibitors, Organic Chemistry, thymidine monophosphate kinase, biology.organism_classification, TMK, MESH: Antitubercular Agents, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, chemistry, DISCOVERY, CHIMERA, Thymine derivatives, Nucleoside-Phosphate Kinase, Nucleoside, MESH: Nucleoside-Phosphate Kinase, SYSTEM
Abstract: We report the design and synthesis of a series of non-nucleoside MtbTMPK inhibitors (1-14) based on the gram-positive bacterial TMPK inhibitor hit compound 1. A practical synthesis was developed to access these analogues. Several compounds show promising MtbTMPK inhibitory potency and allow the establishment of a structure-activity relationship, which is helpful for further optimization. (C) 2016 Elsevier Ltd. All rights reserved.
Published: 2016

36. The Dinoflagellate Toxin 20-Methyl Spirolide-G Potently Blocks Skeletal Muscle and Neuronal Nicotinic Acetylcholine Receptors

Author: Morgane Reynaud, Evelyne Benoit, Denis Servent, Aurélie Couesnon, Rómulo Aráoz, Bogdan I. Iorga, Jordi Molgó, Institut des Neurosciences Paris-Saclay (NeuroPSI), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Institut de Chimie des Substances Naturelles (ICSN), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), This work was supported by the Agence Nationale de la Recherche (France) by Grant ANR-12-ASTR-0037-AQUANEUROTOX., and ANR-12-ASTR-0037,AquaNeuroTox,Identification et caractérisation du mode d'interaction de phycotoxines neurotoxiques présentant un risque biologique - Développement de nouveaux systèmes de détection.(2012)
Subjects: 0301 basic medicine, MESH: Muscle Cells, dinoflagellate toxin, spirolides, nicotinic acetylcholine receptors, neuromuscular transmission, Xenopus oocytes, nicotinic currents, competition-binding assays, molecular docking, MESH: Neurotoxicity Syndromes, Protein Conformation, Pyridines, Health, Toxicology and Mutagenesis, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Neuromuscular transmission, lcsh:Medicine, MESH: Cricetinae, Action Potentials, Stimulation, Nicotinic Antagonists, Receptors, Nicotinic, Toxicology, Torpedo, law.invention, Mice, Xenopus laevis, 0302 clinical medicine, MESH: Structure-Activity Relationship, MESH: Cricetulus, law, MESH: Animals, MESH: Xenopus, MESH: Isometric Contraction, Nicotinic Antagonist, Molecular Docking Simulation, medicine.anatomical_structure, Nicotinic agonist, Biochemistry, Cholinergic Fibers, Epibatidine, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH: Receptors, Nicotinic, Female, medicine.drug, Protein Binding, Neuromuscular Junction, Biology, In Vitro Techniques, Transfection, Binding, Competitive, Neuromuscular junction, Article, MESH: Oocytes, MESH: alpha7 Nicotinic Acetylcholine Receptor, 03 medical and health sciences, Structure-Activity Relationship, MESH: CHO Cells, Isometric Contraction, MESH: Patch-Clamp Techniques, medicine, MESH: Molecular Docking Simulation, Animals, Humans, Spiro Compounds, Muscle, Skeletal, MESH: Mice, Acetylcholine receptor, MESH: Humans, Binding Sites, Dose-Response Relationship, Drug, lcsh:R, MESH: Marine Toxins, Bridged Bicyclo Compounds, Heterocyclic, Molecular biology, Electric Stimulation, MESH: Spiro Compounds, 030104 developmental biology, HEK293 Cells, MESH: Receptors, Muscarinic, MESH: Neuromuscular Junction, Chickens, 030217 neurology & neurosurgery
Abstract: International audience; The cyclic imine toxin 20-methyl spirolide G (20-meSPX-G), produced by the toxigenic dinoflagellate Alexandrium ostenfeldii/Alexandrium peruvianum, has been previously reported to contaminate shellfish in various European coastal locations, as revealed by mouse toxicity bioassay. The aim of the present study was to determine its toxicological profile and its molecular target selectivity. 20-meSPX-G blocked nerve-evoked isometric contractions in isolated mouse neuromuscular preparations, while it had no action on contractions elicited by direct electrical stimulation, and reduced reversibly nerve-evoked compound muscle action potential amplitudes in anesthetized mice. Voltage-clamp recordings in Xenopus oocytes revealed that 20-meSPX-G potently inhibited currents evoked by ACh on Torpedo muscle-type and human α7 nicotinic acetylcholine receptors (nAChR), whereas lower potency was observed in human α4β2 nAChR. Competition-binding assays showed that 20-meSPX-G fully displaced [³H]epibatidine binding to HEK-293 cells expressing the human α3β2 (Ki = 0.040 nM), whereas a 90-fold lower affinity was detected in human α4β2 nAChR. The spirolide displaced [(125)I]α-bungarotoxin binding to Torpedo membranes (Ki = 0.028 nM) and in HEK-293 cells expressing chick chimeric α7-5HT₃ nAChR (Ki = 0.11 nM). In conclusion, this is the first study to demonstrate that 20-meSPX-G is a potent antagonist of nAChRs, and its subtype selectivity is discussed on the basis of molecular docking models.
Published: 2016

37. A novel selective metabotropic glutamate receptor 4 agonist reveals new possibilities for developing subtype selective ligands with therapeutic potential

Author: Delphine Rigault, Francine Acher, Nadia Oueslati, Hervé Daniel, Marianne Amalric, Isabelle Brabet, Jean-Philippe Pin, Heather McLean, Hugues-Olivier Bertrand, Tiphanie Courtiol, Bruno Vilar, Thomas Bessiron, Cyril Goudet, Thierry Deltheil, Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Neurosciences Cognitives [Marseille] (LNC), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Institut de biochimie et biophysique moléculaire et cellulaire (IBBMC), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Accelrys SARL, Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS) - Université Montpellier 2 - Sciences et Techniques (UM2) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Université Montpellier 1 (UM1) - Université de Montpellier (UM), Université Paris Descartes - Paris 5 (UPD5) - Centre National de la Recherche Scientifique (CNRS), Aix Marseille Université (AMU) - Centre National de la Recherche Scientifique (CNRS), and Université Paris-Sud - Paris 11 (UP11) - Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, Patch-Clamp Techniques, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Antiparkinson Agents, [SDV.NEU.PC] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, MESH : Dose-Response Relationship, Drug, Pharmacology, Ligands, Receptors, Metabotropic Glutamate, Synaptic Transmission, Biochemistry, MESH: Mice, Knockout, MESH: Dose-Response Relationship, Drug, Antiparkinson Agents, MESH: Recombinant Proteins, Mice, 0302 clinical medicine, MESH: Structure-Activity Relationship, MESH : Structure-Activity Relationship, Excitatory Amino Acid Agonists, MESH: Ligands, MESH: Animals, MESH : Patch-Clamp Techniques, Receptor, Mice, Knockout, MESH : Synaptic Transmission, MESH : Ligands, 0303 health sciences, Molecular Structure, [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, MESH : Rats, Chemistry, Metabotropic glutamate receptor 4, MESH : Receptors, Metabotropic Glutamate, Glutamate receptor, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, Glutamate binding, MESH : Antiparkinson Agents, MESH : Phosphinic Acids, Recombinant Proteins, MESH : Mutagenesis, Site-Directed, MESH: Mutagenesis, Site-Directed, MESH: HEK293 Cells, Metabotropic glutamate receptor 1, MESH: Phosphinic Acids, Biotechnology, Agonist, MESH: Rats, MESH : Recombinant Proteins, medicine.drug_class, MESH : Male, MESH: Molecular Structure, MESH : Rats, Wistar, Structure-Activity Relationship, 03 medical and health sciences, MESH : Mice, MESH: Patch-Clamp Techniques, Genetics, medicine, MESH: Synaptic Transmission, Animals, Humans, Rats, Wistar, MESH: Receptors, Metabotropic Glutamate, Molecular Biology, MESH: Mice, 030304 developmental biology, MESH : Excitatory Amino Acid Agonists, Binding Sites, MESH: Humans, Dose-Response Relationship, Drug, MESH : Humans, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Rats, Wistar, Phosphinic Acids, MESH: Male, Rats, MESH : HEK293 Cells, HEK293 Cells, Metabotropic receptor, MESH: Binding Sites, Metabotropic glutamate receptor, Mutagenesis, Site-Directed, MESH : Mice, Knockout, MESH : Animals, MESH: Excitatory Amino Acid Agonists, MESH : Molecular Structure, MESH : Binding Sites, [SDV.NEU.SC] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, 030217 neurology & neurosurgery
Abstract: International audience; Metabotropic glutamate (mGlu) receptors are promising targets to treat numerous brain disorders. So far, allosteric modulators are the only subtype selective ligands, but pure agonists still have strong therapeutic potential. Here, we aimed at investigating the possibility of developing subtype-selective agonists by extending the glutamate-like structure to hit a nonconsensus binding area. We report the properties of the first mGlu4-selective orthosteric agonist, derived from a virtual screening hit, LSP4-2022 using cell-based assays with recombinant mGlu receptors [EC(50): 0.11 ± 0.02, 11.6 ± 1.9, 29.2 ± 4.2 μM (n>19) in calcium assays on mGlu4, mGlu7, and mGlu8 receptors, respectively, with no activity at the group I and -II mGlu receptors at 100 μM]. LSP4-2022 inhibits neurotransmission in cerebellar slices from wild-type but not mGlu4 receptor-knockout mice. In vivo, it possesses antiparkinsonian properties after central or systemic administration in a haloperidol-induced catalepsy test, revealing its ability to cross the blood-brain barrier. Site-directed mutagenesis and molecular modeling was used to identify the LSP4-2022 binding site, revealing interaction with both the glutamate binding site and a variable pocket responsible for selectivity. These data reveal new approaches for developing selective, hydrophilic, and brain-penetrant mGlu receptor agonists, offering new possibilities to design original bioactive compounds with therapeutic potential.
Published: 2016

38. Progress with peptide scanning to study structure-activity relationships: the implications for drug discovery

Author: Pierre Tufféry, Stéphanie Eustache, Jérôme Leprince, Enzymologie de l'ARN, Université Pierre et Marie Curie - Paris 6 (UPMC), UMR-S973, MTi, Université Paris Diderot - Paris 7 (UPD7), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Modélisation mathématique et statistique en biologie et médecine, and Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: 0301 basic medicine, MESH: Sequence Analysis, DNA, MESH: Amino Acids, Aza-amino, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Peptide, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, MESH: Drug Design, MESH: Reproduction, MESH: Structure-Activity Relationship, Egg-laying, Drug Discovery, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Amino Acids, chemistry.chemical_classification, Drug discovery, MESH: Peptides, N-alkyl scanning, Pro, MESH: Sex Attractants, Biochemistry, gamma-lactam, D, Functional role, Computational biology, Sepia officinalis, Cephalopod, 03 medical and health sciences, Structure-Activity Relationship, MESH: Gene Expression Profiling, MESH: Drug Discovery, MESH: Gene Library, Humans, MESH: Tissue Distribution, Peptide scanning, MESH: Mass Spectrometry, MESH: Molecular Sequence Data, MESH: Humans, CCAPs, Neuropeptides, NMR, Ala, 030104 developmental biology, chemistry, Drug Design, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Peptide drug, Peptides, Peptide drug development, MESH: Seasons, Positional
Abstract: Peptides have gained renewed interest as candidate therapeutics. However, to bring them to a broader clinical use, challenges such as the rational optimization of their pharmacological properties remain. Peptide scanning techniques offer a systematic framework to gain information on the functional role of individual amino acids of a peptide. Due to progress in mastering new chemical synthesis routes targeting amino acid backbone, they are currently diversified. Structure-activity relationship (SAR) analyses such as alanine- or enantioneric- scanning can now be supplemented by N-substitution, lactam cyclisation- or aza-amino scanning procedures addressing not only SAR considerations but also the peptide pharmacological properties.This review highlights the different scanning techniques currently available and illustrates how they can impact drug discovery.Progress in peptide scanning techniques opens new perspectives for peptide drug development. It comes with the promise of a paradigm change in peptide drug design in which peptide drugs will be closer to the parent peptides. However, scanning still remains assimilable to a trial and error strategy that could benefit from being combined with specific in silico approaches that start reaching maturity.
Published: 2016

39. New imidazoquinoxaline derivatives: Synthesis, biological evaluation on melanoma, effect on tubulin polymerization and structure–activity relationships

Author: Nicole Bec, Jean-François Guichou, Georges Moarbess, Carine Deleuze-Masquéfa, Christian Larroque, Stéphanie Paniagua-Gayraud, Zahraa Zghaib, Mona Diab-Assaf, Pierre-Antoine Bonnet, Issam Kassab, Kamel Hadj-Kaddour, Pierre Cuq, Laure-Anaïs Vincent, Johanna Vappiani, Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM), Lebanese University [Beirut] (LU), Centre de Biochimie Structurale [Montpellier] (CBS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherche en Cancérologie de Montpellier (IRCM - U1194 Inserm - UM), and CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)
Subjects: 0301 basic medicine, Tubulin depolymerization, Clinical Biochemistry, Pharmaceutical Science, Structure–activity relationship, Antiproliferative activity, MESH: Tubulin, Bioinformatics, 01 natural sciences, Biochemistry, Polymerization, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Tubulin, Drug Discovery, Tumor Cells, Cultured, Melanoma, Imidazo[1, Molecular Structure, biology, Chemistry, Human melanoma cancer cell line (A375), MESH: Drug Screening Assays, Antitumor, Imidazoles, 3. Good health, Molecular Docking Simulation, MESH: Polymerization, Molecular docking, Molecular Medicine, MESH: Imidazoles, MESH: Melanoma, MESH: Molecular Structure, Antineoplastic Agents, [SDV.CAN]Life Sciences [q-bio]/Cancer, macromolecular substances, Structure-Activity Relationship, 03 medical and health sciences, Quinoxaline, MESH: Quinoxalines, Microtubule, Quinoxalines, medicine, MESH: Molecular Docking Simulation, Humans, MESH: Tumor Cells, Cultured, Molecular Biology, IC50, MESH: Humans, Dose-Response Relationship, Drug, 010405 organic chemistry, Organic Chemistry, medicine.disease, 2-a]quinoxaline, In vitro, 0104 chemical sciences, 030104 developmental biology, Cell culture, biology.protein, MESH: Antineoplastic Agents, Drug Screening Assays, Antitumor, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: International audience; Microtubules are considered as important targets of anticancer therapy. EAPB0503 and its structural imidazo[1,2-a]quinoxaline derivatives are major microtubule-interfering agents with potent anticancer activity. In this study, the synthesis of several new derivatives of EAPB0503 is described, and the anticancer efficacy of 13 novel derivatives on A375 human melanoma cell line is reported. All new compounds show significant antiproliferative activity with IC50 in the range of 0.077-122μM against human melanoma cell line (A375). Direct inhibition of tubulin polymerization assay in vitro is also assessed. Results show that compounds 6b, 6e, 6g, and EAPB0503 highly inhibit tubulin polymerization with percentages of inhibition of 99%, 98%, 90%, and 84% respectively. Structure-activity relationship studies within the series are also discussed in line with molecular docking studies into the colchicine-binding site of tubulin.
Published: 2016

40. Anti-mycobacterial activity of thymine derivatives bearing boron clusters

Author: Edyta Paradowska, Anna Ruszczyńska, Agnieszka Jabłońska, Ewa Bulska, Anna Adamska, Agnieszka B. Olejniczak, Zbigniew J. Leśnikowski, Mirosława Studzińska, Anna Rumijowska-Galewicz, Jarosław Dziadek, Hélène Munier-Lehmann, Polish Academy of Sciences (PAN), University of Warsaw (UW), Warsaw University of Technology [Warsaw], Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], This work was supported by the following grants: the European Regional Development Fund under the Operational Programme Innovative Economy, grant POIG.01.01.02-10-107/09, and the Statutory Fund of IMB PAS to A. Adamska, A. Rumijowska-Galewicz, M. Studzińska, A. Jabłońska, E. Paradowska, J. Dziadek, Z. J. Leśnikowski, and A. B. Olejniczak. Hélene Munier-Lehmann was supported by CNRS, INSERM and the Institute Pasteur. The ICP MS study was carried out at the Biological and Chemical Research Centre, University of Warsaw, established within the project co-financed by European Union from the European Regional Development Found under Operational Programme Innovative Economy, 2007–2013., The authors thank Patrycja Król, Mateusz Pawlaczyk, and Dominika Staniszewska for assistance with the lipophilicity measurements., and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Boron Compounds, MESH: Thymine, Stereochemistry, [SDV]Life Sciences [q-bio], MESH: Boron Compounds, 010402 general chemistry, 01 natural sciences, Thymidylate kinase, Nucleobase, Mycobacterium, Mycobacterium tuberculosis, chemistry.chemical_compound, Structure-Activity Relationship, MESH: Structure-Activity Relationship, Bacterial Proteins, Anti-mycobacterial activity, MESH: Anti-Bacterial Agents, Drug Discovery, Borates, MESH: Mycobacterium, Structure–activity relationship, [CHIM]Chemical Sciences, Nucleoside, MESH: Bacterial Proteins, MESH: Borates, Pharmacology, biology, 010405 organic chemistry, Mycobacterium smegmatis, Organic Chemistry, MESH: Hydrophobic and Hydrophilic Interactions, General Medicine, biology.organism_classification, 3. Good health, 0104 chemical sciences, Thymine, Anti-Bacterial Agents, Boron cluster, chemistry, Nucleoside-Phosphate Kinase, MESH: Nucleoside-Phosphate Kinase, Hydrophobic and Hydrophilic Interactions
Abstract: International audience; A series of novel thymine derivatives bearing lipophilic, electron-neutral 1,2-dicarba-closo-dodecaborane, 1,12-dicarba-closo-dodecaborane or hydrophilic 7,8-dicarba-nido-undecaborate anions were synthesized. Synthesis was performed via copper(I)-catalysed Huisgen-Meldal-Sharpless 1,3-dipolar cycloaddition of N(1)-propargylthymine or N(1),N(3)-bispropargylthymine to 1-(3-azidopropyl)-1,2-dicarba-closo-dodecaborane. The obtained compounds were tested in vitro against Mycobacterium tuberculosis thymidylate kinase (TMPKmt) and as inhibitors of mycobacteria growth in culture using both saprophytic Mycobacterium smegmatis (M. smegmatis) and pathogenic Mycobacterium tuberculosis (M. tuberculosis) strains. The most potent TMPKmt inhibitor in the series contained two negatively charged 7,8-dicarba-nido-undecaborate modifications at positions 1 and 3 of thymine (9) and exhibited a Ki value of 1.5 μM. The most potent inhibitors of mycobacteria growth was compound 5 with one electron-neutral 1,2-dicarba-closo-dodecaborane modification at position 1 of thymine, and compound 8 with two modifications, at position 1 and 3. Both compounds completely inhibited M. smegmatis proliferation at a concentration of 100 μg/mL (0.25 mM and 0.15 mM, respectively).
Published: 2016

41. Structure-Activity Relationship Studies of N- and C-Terminally Modified Secretin Analogs for the Human Secretin Receptor

Author: Billy K. C. Chow, Jérôme Leprince, Kailash Singh, Ahmed Aa Allam, Jamaan S. Ajarem, Aloysius Wilfred Raj Arokiaraj, Vijayalakshmi Senthil, David Vaudry, Benjamin Lefranc, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), The University of Hong Kong (HKU), King Saud University [Riyadh] (KSU), and Beni-Suef University
Subjects: 0301 basic medicine, Peptide Hormones, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, lcsh:Medicine, Plasma protein binding, MESH: Amino Acid Sequence, MESH: Receptors, G-Protein-Coupled, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Biochemistry, Fluorophotometry, Receptors, G-Protein-Coupled, Secretin, Binding Analysis, Spectrum Analysis Techniques, 0302 clinical medicine, MESH: Structure-Activity Relationship, Drug Discovery, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Cyclic AMP, Fluorescence Resonance Energy Transfer, Post-Translational Modification, lcsh:Science, Peptide sequence, MESH: Cyclic AMP, ComputingMilieux_MISCELLANEOUS, MESH: Secretin, 0303 health sciences, Crystallography, Multidisciplinary, MESH: Peptides, Physics, Condensed Matter Physics, 3. Good health, Molecular Docking Simulation, Spectrophotometry, 030220 oncology & carcinogenesis, Physical Sciences, Crystal Structure, Sequence Analysis, Signal Peptides, Protein Binding, Research Article, Signal Transduction, Agonist, Transmembrane Receptors, medicine.drug_class, Molecular Sequence Data, Allosteric regulation, Secretin receptor family, Biology, Research and Analysis Methods, Receptors, Gastrointestinal Hormone, Structure-Activity Relationship, 03 medical and health sciences, MESH: Drug Discovery, medicine, MESH: Molecular Docking Simulation, Humans, Solid State Physics, MESH: Protein Binding, Amino Acid Sequence, Molecular Biology Techniques, Sequencing Techniques, Molecular Biology, Chemical Characterization, 030304 developmental biology, G protein-coupled receptor, MESH: Humans, MESH: Molecular Sequence Data, 030102 biochemistry & molecular biology, MESH: Receptors, Gastrointestinal Hormone, lcsh:R, Correction, Biology and Life Sciences, Proteins, Cell Biology, Hormones, 030104 developmental biology, Docking (molecular), [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, lcsh:Q, G Protein Coupled Receptors, Peptides, Sequence Alignment
Abstract: The pleiotropic role of human secretin (hSCT) validates its potential use as a therapeutic agent. Nevertheless, the structure of secretin in complex with its receptor is necessary to develop a suitable therapeutic agent. Therefore, in an effort to design a three-dimensional virtual homology model and identify a peptide agonist and/or antagonist for the human secretin receptor (hSR), the significance of the primary sequence of secretin peptides in allosteric binding and activation was elucidated using virtual docking, FRET competitive binding and assessment of the cAMP response. Secretin analogs containing various N- or C-terminal modifications were prepared based on previous findings of the role of these domains in receptor binding and activation. These analogs exhibited very low or no binding affinity in a virtual model, and were found to neither exhibit in vitro binding nor agonistic or antagonistic properties. A parallel analysis of the analogs in the virtual model and in vitro studies revealed instability of these peptide analogs to bind and activate the receptor., published_or_final_version
Published: 2016

42. Influenza Polymerase Can Adopt an Alternative Configuration Involving a Radical Repacking of PB2 Domains

Author: Alexander Pflug, Thomas Bock, Narin Hengrung, Delphine Guilligay, Darren J. Hart, Kamel El Omari, Florence Baudin, Eric Thierry, Stephen Cusack, Stephanie Gaudon, Martin Beck, Jan Kosinski, Adam Round, European Molecular Biology Laboratory [Grenoble] (EMBL), European Molecular Biology Laboratory [Heidelberg] (EMBL), Division of Structural Biology, University of Oxford [Oxford], Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), University of Oxford, Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Subjects: 0301 basic medicine, Models, Molecular, Influenzavirus C, viruses, Nuclear Localization Signals, MESH: Nuclear Localization Signals, MESH: Amino Acid Sequence, Crystallography, X-Ray, Mass Spectrometry, MESH: Influenzavirus C, MESH: Structure-Activity Relationship, Transcription (biology), MESH: Influenza B virus, Polymerase, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], 3. Good health, Ribonucleoproteins, MESH: RNA, Viral, RNA, Viral, MESH: Lasers, Transcription factor II D, MESH: RNA Replicase, MESH: Models, Molecular, MESH: Influenza A Virus, H5N1 Subtype, Molecular Sequence Data, RNA-dependent RNA polymerase, Article, Cap snatching, 03 medical and health sciences, Structure-Activity Relationship, Viral Proteins, Scattering, Small Angle, RNA polymerase I, Humans, Protein Interaction Domains and Motifs, Amino Acid Sequence, Molecular Biology, MESH: Scattering, Small Angle, MESH: Mass Spectrometry, Messenger RNA, MESH: Protein Interaction Domains and Motifs, MESH: Humans, MESH: Molecular Sequence Data, Influenza A Virus, H5N1 Subtype, Lasers, RNA, Cell Biology, biochemical phenomena, metabolism, and nutrition, RNA-Dependent RNA Polymerase, MESH: Crystallography, X-Ray, Molecular biology, MESH: Viral Proteins, Influenza B virus, MESH: Ribonucleoproteins, 030104 developmental biology, biology.protein
Abstract: Summary Influenza virus polymerase transcribes or replicates the segmented RNA genome (vRNA) into respectively viral mRNA or full-length copies and initiates RNA synthesis by binding the conserved 3′ and 5′ vRNA ends (the promoter). In recent structures of promoter-bound polymerase, the cap-binding and endonuclease domains are configured for cap snatching, which generates capped transcription primers. Here, we present a FluB polymerase structure with a bound complementary cRNA 5′ end that exhibits a major rearrangement of the subdomains within the C-terminal two-thirds of PB2 (PB2-C). Notably, the PB2 nuclear localization signal (NLS)-containing domain translocates ∼90 Å to bind to the endonuclease domain. FluA PB2-C alone and RNA-free FluC polymerase are similarly arranged. Biophysical and cap-dependent endonuclease assays show that in solution the polymerase explores different conformational distributions depending on which RNA is bound. The inherent flexibility of the polymerase allows it to adopt alternative conformations that are likely important during polymerase maturation into active progeny RNPs., Graphical Abstract, Highlights • Flexibly linked influenza polymerase domains found to pack in a new configuration • PB2 nuclear localization signal domain translocates 93 Å to pack on PA endonuclease • Similar arrangement observed in influenza A, B, and C polymerases • Multiple conformations accessible in solution depending on which viral RNA is bound, Influenza polymerase is a multifunctional machine that both replicates and transcribes the viral RNA genome. Thierry et al. show that different functional states can occur by repacking of peripheral domains flexibly linked to the polymerase core depending on which kind of viral RNA is bound.
Published: 2016

43. eIF3 Peripheral Subunits Rearrangement after mRNA Binding and Start-Codon Recognition

Author: Adeline Renaud, A. Myasnikov, Lauriane Kuhn, Angelita Simonetti, Yaser Hashem, Jailson Brito Querido, Eder Mancera-Martínez, MATHELIN, Sandra, Initiative d'excellence - Par-delà les frontières, l'Université de Strasbourg - - UNISTRA2010 - ANR-10-IDEX-0002 - IDEX - VALID, Architecture et Réactivité de l'ARN (ARN), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Plateforme Protéomique Strasbourg - Esplanade (IBMC / CNRS FRC1589 / UNIV Strasbourg), Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Moléculaire et Cellulaire [Strasbourg] (IBMC), This work was supported by the ANR grant @RAction program ‘‘ANR CryoEM80S’’ and the LABEX: ANR-10-LABX-0036_NETRNA and benefits from funding from the state managed by the French National Research Agency as part of the Investments for the Future program (to Y.H.)., ANR-10-IDEX-0002,UNISTRA,Par-delà les frontières, l'Université de Strasbourg(2010), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)
Subjects: Models, Molecular, 0301 basic medicine, MESH: Eukaryotic Initiation Factor-3, Five prime untranslated region, MESH: Eukaryotic Initiation Factor-2, Protein Conformation, Eukaryotic Initiation Factor-3, MESH: Codon, Initiator, [SDV]Life Sciences [q-bio], Eukaryotic Initiation Factor-2, Eukaryotic Initiation Factor-1, Codon, Initiator, MESH: Rabbits, beta-Globins, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Structure-Activity Relationship, MESH: Protein Conformation, RNA, Transfer, Eukaryotic initiation factor, MESH: Animals, MESH: Protein Subunits, [SDV] Life Sciences [q-bio], MESH: Nucleic Acid Conformation, ribosome, MESH: Protein Biosynthesis, Transfer RNA, Rabbits, eIF3, MESH: Eukaryotic Initiation Factor-1, MESH: Models, Molecular, Protein Binding, eukaryotic translation initiation, Biology, Structure-Activity Relationship, 03 medical and health sciences, Eukaryotic translation, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, Humans, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, RNA, Messenger, Molecular Biology, eIF3 peripheral subunits, MESH: RNA, Messenger, Binding Sites, MESH: Humans, MESH: beta-Globins, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Biology, MESH: Multiprotein Complexes, MESH: RNA, Transfer, Molecular biology, Ribosomal binding site, Protein Subunits, Internal ribosome entry site, 030104 developmental biology, MESH: Binding Sites, Multiprotein Complexes, Protein Biosynthesis, eIF4A, Nucleic Acid Conformation, cryo-EM, Ribosomes, MESH: Ribosomes
Abstract: International audience; mRNA translation initiation in eukaryotes requires the cooperation of a dozen eukaryotic initiation factors (eIFs) forming several complexes, which leads to mRNA attachment to the small ribosomal 40S subunit, mRNA scanning for start codon, and accommodation of initiator tRNA at the 40S P site. eIF3, composed of 13 subunits, 8 core (a, c, e, f, h, l, k, and m) and 5 peripheral (b, d, g, i, and j), plays a central role during this process. Here we report a cryo-electron microscopy structure of a mammalian 48S initiation complex at 5.8 Å resolution. It shows the relocation of subunits eIF3i and eIF3g to the 40S intersubunit face on the GTPase binding site, at a late stage in initiation. On the basis of a previous study, we demonstrate the relocation of eIF3b to the 40S intersubunit face, binding below the eIF2-Met-tRNAi(Met) ternary complex upon mRNA attachment. Our analysis reveals the deep rearrangement of eIF3 and unravels the molecular mechanism underlying eIF3 function in mRNA scanning and timing of ribosomal subunit joining.
Published: 2016

44. Design, Synthesis, and Structure–Activity Relationship of N-Arylnaphthylamine Derivatives as Amyloid Aggregation Inhibitors

Author: Luca Pescatori, Orlando Ghirardi, Roberto Di Santo, Diana Celona, Giovanna Guiso, Federica Rosi, Fabrizio Giorgi, Roberta Costi, Mario Vertechy, Patrizia Minetti, Paola Piovesan, Luigi Scipione, Mauro Marzi, Giuliana Cuzzucoli Crucitti, Silvio Caccia, Department of Medicinal Chemistry and Technologies, Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Sigma-Tau S.p.A., Pomezia, Dipartimento di Neuroscienze, and Istituto di Ricerche Farmacologiche 'Mario Negri'
Subjects: Amyloid, Stereochemistry, Naphthalenes, MESH: Drug Design, Fibril, MESH: Blood-Brain Barrier, Mice, Structure-Activity Relationship, 03 medical and health sciences, MESH: Structure-Activity Relationship, 0302 clinical medicine, Drug Discovery, medicine, Animals, Structure–activity relationship, Potency, Tissue Distribution, MESH: Animals, MESH: Tissue Distribution, MESH: Peptide Fragments, MESH: Mice, 030304 developmental biology, MESH: Amyloid, 0303 health sciences, Amyloid beta-Peptides, Chemistry, MESH: Naphthalenes, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, medicine.disease, Peptide Fragments, MESH: Amyloid beta-Peptides, In vitro, 3. Good health, Design synthesis, Biochemistry, Blood-Brain Barrier, Drug Design, Amyloid aggregation, Molecular Medicine, Alzheimer's disease, 030217 neurology & neurosurgery
Abstract: International audience; Dyes like CR are able to inhibit the aggregation of Aβ fibrils. Thus, a screening of a series of dyes including ABBB (1) was performed. Its main component 2 tested in an in vitro assay (i.e., ThT assay) showed good potency at inhibiting fibrils association. Congeners 4-9 have been designed and synthesized as inhibitors of Aβ aggregation. A number of these newly synthesized compounds have been found to be active in the ThT assay with IC(50) of 1-57.4 μM. The most potent compound of this series, 4k, showed micromolar activity in this test. Another potent derivative 4q (IC(50) = 5.6 μM) rapidly crossed the blood-brain barrier, achieving whole brain concentrations higher than in plasma. So 4q could be developed to find novel potent antiaggregating βA agents useful in Alzheimer disease as well as other neurological diseases characterized by deposits of amyloid aggregates.
Published: 2012

45. Structure–activity relationship of selected polyphenol derivatives as inhibitors of Bax/Bcl-xL interaction

Author: René Grée, Fabien Gautier, Philippe Juin, Duc Duy Vo, Institut des Sciences Chimiques de Rennes (ISCR), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA), Centre de Recherche en Cancérologie Nantes-Angers (CRCNA), Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM)-Hôtel-Dieu de Nantes-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital Laennec-Centre National de la Recherche Scientifique (CNRS)-Faculté de Médecine d'Angers-Centre hospitalier universitaire de Nantes (CHU Nantes), Ligue contre le Cancer, Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Stereochemistry, bcl-X Protein, Antineoplastic Agents, Apoptosis, Bcl-xL, MESH: bcl-X Protein, Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, MESH: Structure-Activity Relationship, 0302 clinical medicine, Drug Discovery, MESH: Protein Binding, Humans, Phenol, Structure–activity relationship, MESH: bcl-2-Associated X Protein, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], bcl-2-Associated X Protein, 030304 developmental biology, Pharmacology, 0303 health sciences, MESH: Humans, biology, MESH: Apoptosis, Organic Chemistry, Polyphenols, General Medicine, 3. Good health, chemistry, Biochemistry, Polyphenol, 030220 oncology & carcinogenesis, MESH: HeLa Cells, biology.protein, MESH: Antineoplastic Agents, MESH: Polyphenols, HeLa Cells, Protein Binding
Abstract: International audience; This paper describes the synthesis of nine selected diaryl/heteroaryl-containing phenol and polyphenol derivatives which have been evaluated against Bax/Bcl-xL interaction in comparison with ABT-737. Using a BRET assay, six of these derivatives exhibit activity comparable to ABT-737 to disrupt Bax/Bcl-xL interaction. These preliminary results demonstrate that such polyphenol-derived molecules are attractive compounds regarding anticancer activity and that the phenol at position 3 is important regarding disruption of Bax/Bcl-xL interaction.
Published: 2012

46. Synthesis of chromeno[3,4-b]indoles as Lamellarin D analogues : A novel DYRK1A inhibitor class

Author: Frédéric Buron, Emeline Vedrenne, Cleopatra Neagoie, Olivier Lozach, Laurent Meijer, Stéphane Bourg, Sorin Rosca, Amélie Lansiaux, Sylvain Routier, Brigitte Baldeyrou, Jean-Yves Mérour, Institut de Chimie Organique et Analytique (ICOA), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Universitatea Politehnica din Bucuresti [Bucarest, Roumanie] (UPB), Synthèse Organique et Réactivité (SOR), Unité de Chimie et Procédés (UCP), École Nationale Supérieure de Techniques Avancées (ENSTA Paris)-École Nationale Supérieure de Techniques Avancées (ENSTA Paris), Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre Régional de Lutte contre le Cancer Oscar Lambret [Lille] (UNICANCER/Lille), Université de Lille-UNICANCER, Molécules et cibles thérapeutiques (MCT), Station biologique de Roscoff [Roscoff] (SBR), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U837 (JPArc), Université Lille Nord de France (COMUE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Université d'Orléans (UO)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Université Lille Nord de France (COMUE)-UNICANCER, Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U1172 Inserm - U837 (JPArc), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université d'Orléans (UO)-Institut de Chimie du CNRS (INC), ManRos Therapeutics, and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE)-Université de Lille
Subjects: Models, Molecular, Indoles, DYRK1A, Stereochemistry, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Protein Serine-Threonine Kinases, 010402 general chemistry, Heterocyclic Compounds, 4 or More Rings, MESH: Protein-Tyrosine Kinases, 01 natural sciences, MESH: Protein-Serine-Threonine Kinases, Structure-Activity Relationship, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Coumarins, Drug Discovery, MESH: Isoquinolines, Humans, MESH: Protein Kinase Inhibitors, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Protein Kinase Inhibitors, ComputingMilieux_MISCELLANEOUS, MESH: Heterocyclic Compounds with 4 or More Rings, MESH: Indoles, Pharmacology, Indole test, MESH: Humans, biology, 010405 organic chemistry, Chemistry, Kinase, Topoisomerase, Organic Chemistry, Halogenation, MESH: Coumarins, General Medicine, Protein-Tyrosine Kinases, Isoquinolines, 3. Good health, 0104 chemical sciences, MESH: Topoisomerase I Inhibitors, DNA Topoisomerases, Type I, biology.protein, Lamellarin D, Topoisomerase I Inhibitors, Pharmacophore, MESH: DNA Topoisomerases, Type I, MESH: Models, Molecular, Derivative (chemistry)
Abstract: International audience; A library of substituted chromeno[3,4-b]indoles was developed as Lamellarin isosters. Synthesis was achieved from indoles after a four-step pathway sequence involving C-3 iodination, a Suzuki cross-coupling reaction, and a one pot deprotection/lactonisation step. Twenty final compounds were tested in order to determine their activity against topoisomerase I and kinases, the two major biological activities of Lamellarins. One newly synthesized derivative exhibited a strong topoisomerase activity comparable to reference compounds such as campthotecin and Lamellarin with only a weak kinase inhibition. Two other lead compounds were identified as new nanomolar DYRK1A inhibitors and several other drugs affected the kinases in the sub-micromolar range. These results will enable us to use the chromeno[3,4-b]indole as a pharmacophore to develop potent treatments for neurological or oncological disorders in which DYRK1A is fully involved.
Published: 2012

47. Novel Reversible Monoamine Oxidase A Inhibitors: Highly Potent and Selective 3-(1H-Pyrrol-3-yl)-2-oxazolidinones

Author: Stefania Saccoccio, Sergio Valente, Stefano Tomassi, Enzo Agostinelli, Antonello Mai, G. Tempera, Department of Medicinal Chemistry and Technologies, Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], and Department of Biochemical Sciences 'Rossi Fanelli'
Subjects: Monoamine Oxidase Inhibitors, Monoamine oxidase, MESH: Monoamine Oxidase Inhibitors, Stereoisomerism, In Vitro Techniques, Pharmacology, MESH: Monoamine Oxidase, 01 natural sciences, Isozyme, Structure-Activity Relationship, 03 medical and health sciences, MESH: Structure-Activity Relationship, MESH: Oxazolidinones, Drug Discovery, medicine, Animals, Structure–activity relationship, Dementia, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Pyrroles, Monoamine Oxidase, Oxazolidinones, 030304 developmental biology, 0303 health sciences, 010405 organic chemistry, Chemistry, MAO inhibitors, medicine.disease, MESH: Stereoisomerism, Monoamine Oxidase-A Inhibitors, 3. Good health, 0104 chemical sciences, Isoenzymes, MESH: Cattle, Monoamine neurotransmitter, MESH: Isoenzymes, MESH: Pyrroles, Molecular Medicine, Cattle
Abstract: Monoamine oxidases (MAOs) are involved in various psychiatric and neurodegenerative disorders; hence, MAO inhibitors are useful agents in the therapy of Parkinson's disease, Alzheimer's dementia, and depression syndrome. Herein we report a novel series of 3-(1H-pyrrol-3-yl)-2-oxazolidinones 3-7 as reversible, highly potent and selective anti-MAO-A agents. In particular, 4b, 5b, and 4c showed a K(i-MAO-A) of 0.6, 0.8, and 1 nM, respectively, 4c being 200000-fold selective for MAO-A with respect to MAO-B.
Published: 2011

48. Diketo Acids Derivatives as Dual Inhibitors of Human Immunodeficiency Virus Type 1 Integrase and the Reverse Transcriptase RNase H Domain

Author: R. Di Santo, Department of Medicinal Chemistry and Technologies, Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, and Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome]
Subjects: DNA polymerase, Quinolones, 01 natural sciences, Biochemistry, chemistry.chemical_compound, quinolinone drivatives, MESH: Structure-Activity Relationship, Drug Discovery, 0303 health sciences, biology, diketo acids, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, Keto Acids, IN inhibitor, HIV Reverse Transcriptase, 3. Good health, Integrase, pyrrole derivatives, RNase H inhibitor, aids, dual inhibitors, hiv, hiv-1, in inhibitor, integrase, reverse transcriptase, ribonuclease h, rnase h, rnase h inhibitor, MESH: Pyrroles, Reverse Transcriptase Inhibitors, Molecular Medicine, RNase H, MESH: HIV Reverse Transcriptase, RNase P, MESH: Keto Acids, Structure-Activity Relationship, 03 medical and health sciences, Humans, Pyrroles, HIV Integrase Inhibitors, MESH: HIV Integrase Inhibitors, 030304 developmental biology, MESH: Ribonuclease H, Pharmacology, MESH: Humans, MESH: Quinolones, Organic Chemistry, RNA, Molecular biology, Reverse transcriptase, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, RNase MRP, chemistry, HIV-1, biology.protein, MESH: Reverse Transcriptase Inhibitors, ribonuclease H, DNA
Abstract: The HIV-1 integrase (IN) and reverse transcriptase (RT) are essential enzymes in the virus cycle. RT is crucial for the retrotranscription of the RNA viral genome, while IN is involved in the insertion in host chromosome of the proviral double strand DNA produced by RT. This enzyme has two associated functions: the RNA- and DNA-dependent DNA polymerase (RDDP and DDDP) and the ribonuclease H (RNase H). The RNase H function catalyzes the selective hydrolysis of the RNA strand of the RNA:DNA heteroduplex replication intermediate. Since the discovery that catalytic cores of both HIV-1 RNase H and IN are folded in a very similar way, have very similar active site geometries, and show the same DDE triad absolutely required for catalytic activity, some researches were devoted to study IN and RNase H dual inhibitor. Our decennial interest in design and synthesis of IN inhibitors led us to study the activity of our compounds also on RNase H activity. The results of the activities showed by pyrrolyl and quinolonyl diketo acids are reported and discussed.
Published: 2011

49. Structure–Activity Relationships of a Series of Analogues of the RFamide-Related Peptide 26RFa

Author: Le Marec, Olivier, Neveu, Cindy, Lefranc, Benjamin, Dubessy, Christophe, Boutin, Jean, Do-Régo, Jean, Costentin, Jean, Tonon, Marie-Christine, Tena-Sempere, Manuel, Vaudry, Hubert, Do-Régo, Jean, Leprince, Jérôme, EPSM-Morbihan, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherches Servier, Centre de Recherches de Croissy, Neuropsycho-pharmacologie expérimentale, Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Department of Cell Biology, and Neuroendocrinologie cellulaire et moléculaire
Subjects: [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Cricetinae, Peptide, MESH: Amino Acid Sequence, MESH: Receptors, G-Protein-Coupled, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Ligands, MESH: Neuropeptides, Receptors, G-Protein-Coupled, MESH: GTP-Binding Protein beta Subunits, chemistry.chemical_compound, MESH: Structure-Activity Relationship, 0302 clinical medicine, MESH: Cricetulus, Cricetinae, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Drug Discovery, MESH: Ligands, MESH: Animals, Peptide sequence, Alanine, chemistry.chemical_classification, 0303 health sciences, GTP-Binding Protein beta Subunits, Stereoisomerism, Gonadotropin secretion, Biochemistry, MESH: Calcium, Molecular Medicine, Stereochemistry, CHO Cells, Transfection, Structure-Activity Relationship, 03 medical and health sciences, Cricetulus, MESH: CHO Cells, Animals, Structure–activity relationship, Amino Acid Sequence, 030304 developmental biology, G protein-coupled receptor, MESH: Transfection, Neuropeptides, Rational design, MESH: Stereoisomerism, chemistry, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Calcium, Norvaline, 030217 neurology & neurosurgery
Abstract: International audience; 26RFa is a new member of the RFamide peptide family that has been identified as the endogenous ligand of the orphan GPCR GPR103. As the C-terminal heptapeptide (26RFa((20-26))) mimics the action of the native peptide on food intake and gonadotropin secretion in rodents, we have synthesized a series of analogues of 26RFa((20-26)) and measured their potency to induce [Ca(2+)](i) mobilization in Gα(16)-hGPR103-transfected CHO cells. Systematic replacement of each residue by an alanine (Ala scan) and its D-enantiomer (D scan) showed that the last three C-terminal residues were very sensitive to the substitutions while position 23 tolerated rather well both modifications. Most importantly, replacement of Ser(23) by a norvaline led to an analogue, [Nva(23)]26RFa((20-26)), that was 3-fold more potent than the native heptapeptide. These new pharmacological data, by providing the first information regarding the structure-activity relationships of 26RFa analogues, should prove useful for the rational design of potent GPR103 receptor ligands with potential therapeutic application.
Published: 2011

50. Structure-Based Design of Short Peptide Ligands Binding onto the E. coli Processivity Ring

Author: Eric Ennifar, Philippe Dumas, Vincent Olieric, Dominique Burnouf, Jean Paul Briand, Jérôme Wagner, Annick Dejaegere, Olivier Chaloin, Gilles Guichard, Philippe Wolff, Architecture et Réactivité de l'ARN (ARN), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), The Swiss Light Source (SLS) (SLS-PSI), Paul Scherrer Institute (PSI), Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Chimie et Biologie des Membranes et des Nanoobjets (CBMN), Université de Bordeaux (UB)-École Nationale d'Ingénieurs des Travaux Agricoles - Bordeaux (ENITAB)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Biotechnologie et signalisation cellulaire (BSC), and Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche de l'Ecole de biotechnologie de Strasbourg (IREBS)
Subjects: Models, Molecular, DNA polymerase, Stereochemistry, MESH: DNA Polymerase III, MESH: Escherichia coli Proteins, MESH: Drug Design, Crystallography, X-Ray, Ligands, Ring (chemistry), Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Sciences du Vivant [q-bio]/Autre [q-bio.OT], MESH: DNA Polymerase beta, Drug Discovery, MESH: Ligands, Escherichia coli, MESH: Protein Binding, DNA Polymerase beta, Peptide ligand, DNA Polymerase III, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, biology, MESH: Escherichia coli, Escherichia coli Proteins, 030302 biochemistry & molecular biology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Processivity, MESH: Crystallography, X-Ray, Ligand (biochemistry), Orders of magnitude (mass), Enzyme, chemistry, Drug Design, MESH: Oligopeptides, biology.protein, Thermodynamics, Molecular Medicine, MESH: Thermodynamics, Oligopeptides, MESH: Models, Molecular, DNA, Protein Binding
Abstract: International audience; The multimeric DNA sliding clamps confer high processivity to replicative DNA polymerases and are also binding platforms for various enzymes involved in DNA metabolism. These enzymes interact with the clamp through a small peptide that binds into a hydrophobic pocket which is a potential target for the development of new antibacterial compounds. Starting from a generic heptapeptide, we used a structure-based strategy to improve the design of new peptide ligands. Chemical modifications at specific residues result in a dramatic increase of the interaction as measured by SPR and ITC. The affinity of our best hits was improved by 2 orders of magnitude as compared to the natural ligand, reaching 10(-8) M range. The molecular basis of the interactions was analyzed by solving the co-crystal structures of the most relevant peptides bound to the clamp and reveals how chemical modifications establish new contacts and contributes to an increased affinity of the ligand.
Published: 2011

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