Search

Your search keyword '"François-Xavier Cantrelle"' showing total 20 results
Start Over Author "François-Xavier Cantrelle" Topic humans
20 results on '"François-Xavier Cantrelle"'

1. Hepatitis E virus RNA‐dependent RNA polymerase is involved in RNA replication and infectious particle production

Author

Jérôme Gouttenoire, Noémie Oechslin, Nathalie Da Silva, François-Xavier Cantrelle, Xavier Hanoulle, Darius Moradpour, Dagmara Szkolnicka, Université de Lausanne = University of Lausanne (UNIL), Biologie Structurale Intégrative (ERL 9002 - BSI ), 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 National de la Recherche Scientifique (CNRS), Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 (RID-AGE), 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), This work was supported by the Swiss National Science Foundation (31003A-179424 to DM and CRSK-3_190706 to JG), the Novartis Foundation (18C140 to DM)as well as the French Agency for Research on AIDS and Viral Hepatitis (ECTZ101316 to XH). The NMR facilities were funded by the North Region Council, CNRS, European Union (FEDER), French Research Ministry and University of Lille., HANOULLE, Xavier, and Université de Lausanne (UNIL)

Subjects
Protein Conformation, alpha-Helical, RdRp, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], viruses, particle production, RNA-dependent RNA polymerase, Biology, Virus Replication, medicine.disease_cause, Genome, Virus, Structure-Activity Relationship, reverse genetics, 03 medical and health sciences, chemistry.chemical_compound, Hepatitis E virus, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, RNA polymerase, medicine, Humans, replicase, 030304 developmental biology, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, 0303 health sciences, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Hepatology, 030306 microbiology, thumb subdomain, RNA, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Hep G2 Cells, RNA-Dependent RNA Polymerase, Virology, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Reverse genetics, Hepatitis E, Open reading frame, chemistry, HEV, Mutation, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, RNA, Viral, replicon
Abstract

International audience; Background and Aims: Hepatitis E virus (HEV) is one of the most common causes of acute hepatitis worldwide. Its positive-strand RNA genome encodes three open reading frames (ORF). ORF1 is translated into a large protein composed of multiple domains and known as the viral replicase. The RNA-dependent RNA polymerase (RdRp) domain is responsible for the synthesis of viral RNA.Approach and Results: Here, we identified a highly conserved α-helix located in the RdRp thumb subdomain. Nuclear magnetic resonance demonstrated an amphipathic α-helix extending from amino acids 1628 to 1644 of the ORF1 protein. Functional analyses revealed a dual role of this helix in HEV RNA replication and virus production, including assembly and release. Mutations on the hydrophobic side of the amphipathic α-helix impaired RNA replication and resulted in the selection of a second-site compensatory change in the RdRp palm subdomain. Other mutations enhanced RNA replication but impaired virus assembly and/or release.Conclusions: Structure-function analyses identified a conserved amphipathic α-helix in the thumb subdomain of the HEV RdRp with a dual role in viral RNA replication and infectious particle production. This study provides structural insights into a key segment of the ORF1 protein and describes the successful use of reverse genetics in HEV, revealing functional interactions between the RdRp thumb and palm subdomains. On a broader scale, it demonstrates that the HEV replicase, similar to those of other positive-strand RNA viruses, is also involved in virus production

Published
2021

2. Dynamic interactions and Ca2+-binding modulate the holdase-type chaperone activity of S100B preventing tau aggregation and seeding

Author

Urmi Sengupta, Guilherme G. Moreira, Ana P. Carapeto, Filipa S. Carvalho, Andrea Quezada, Mário Rodrigues, Isabel Cardoso, Guenter Fritz, Nicha Puangmalai, Isabelle Landrieu, Federico Herrera, Rakez Kayed, Cláudio M. Gomes, Joana S. Cristóvão, François Xavier Cantrelle, Universidade de Lisboa = University of Lisbon (ULISBOA), Biologie Structurale Intégrative (ERL 9002 - BSI ), 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 National de la Recherche Scientifique (CNRS), Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 (RID-AGE), 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), The University of Texas Medical Branch (UTMB), Universidade do Porto = University of Porto, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), University of Hohenheim, This work was funded by Fundação para a Ciência e Tecnologia (Portugal) through research grants PTDC/NEU-NMC/2138/2014 (to C.M.G.), PTDC/BIA-BQM/29963/2017 (F.S.C.), PTDC/MED-NEU/31417/2017 (to F.H.), and POCI-01-0145-FEDER-007274 (to I.C.), investigator grants CEECIND/00031/2017 (to A.P.C.) and IF/00094/2013/CP1173/CT0005 (to F.H.), PhD fellowship SFRH/BD/101171/2014 (to J.S.C.) and DFA/BD/6443/2020 (to G.G.M.), and center grants UIDB/04046/2020 and UID/MULTI/04046/2020 (to BioISI) and Norte-01-0145-FEDER-000008 (to IBMC/I3S)., Landrieu, Isabelle, Universidade de Lisboa (ULISBOA), and Universidade do Porto

Subjects
[SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Science, Tau protein, General Physics and Astronomy, tau Proteins, S100 Calcium Binding Protein beta Subunit, Protein Aggregation, Pathological, Article, Biophysical Phenomena, General Biochemistry, Genetics and Molecular Biology, Cell Line, 03 medical and health sciences, 0302 clinical medicine, Microtubule, Biophysical chemistry, Calcium-binding protein, mental disorders, Humans, Protein folding, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Nuclear Magnetic Resonance, Biomolecular, [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], 030304 developmental biology, 0303 health sciences, Multidisciplinary, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Chemistry, Neurodegenerative Diseases, General Chemistry, Kinetics, Proteostasis, Structural biology, Chaperone (protein), biology.protein, Biophysics, Protein Structural Elements, 030217 neurology & neurosurgery, Molecular Chaperones, Protein Binding, Binding domain
Abstract

The microtubule-associated protein tau is implicated in the formation of oligomers and fibrillar aggregates that evade proteostasis control and spread from cell-to-cell. Tau pathology is accompanied by sustained neuroinflammation and, while the release of alarmin mediators aggravates disease at late stages, early inflammatory responses encompass protective functions. This is the case of the Ca2+-binding S100B protein, an astrocytic alarmin which is augmented in AD and which has been recently implicated as a proteostasis regulator, acting over amyloid β aggregation. Here we report the activity of S100B as a suppressor of tau aggregation and seeding, operating at sub-stoichiometric conditions. We show that S100B interacts with tau in living cells even in microtubule-destabilizing conditions. Structural analysis revealed that tau undergoes dynamic interactions with S100B, in a Ca2+-dependent manner, notably with the aggregation prone repeat segments at the microtubule binding regions. This interaction involves contacts of tau with a cleft formed at the interface of the S100B dimer. Kinetic and mechanistic analysis revealed that S100B inhibits the aggregation of both full-length tau and of the microtubule binding domain, and that this proceeds through effects over primary and secondary nucleation, as confirmed by seeding assays and direct observation of S100B binding to tau oligomers and fibrils. In agreement with a role as an extracellular chaperone and its accumulation near tau positive inclusions, we show that S100B blocks proteopathic tau seeding. Together, our findings establish tau as a client of the S100B chaperone, providing evidence for neuro-protective functions of this inflammatory mediator across different tauopathies., The calcium binding protein S100B is an abundantly expressed protein in the brain and has neuro-protective functions by inhibiting Aβ aggregation and metal ion toxicity. Here, the authors combine cell biology and biochemical experiments with chemical kinetics and NMR measurements and show that S100B protein is an extracellular Tau chaperone and further characterize the interactions between S100B and Tau.

Published
2021

3. Phosphorylated full-length Tau interacts with 14-3-3 proteins via two short phosphorylated sequences, each occupying a binding groove of 14-3-3 dimer

Author

João Filipe Neves, François Xavier Cantrelle, Tomas Obsil, Isabelle Landrieu, Gavin O'Mahony, Xavier Hanoulle, Olivia Petrvalska, Francesco Bosica, Hamida Merzougui, Chemical Biology, Biologie Structurale Intégrative (ERL 9002 - BSI ), 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 National de la Recherche Scientifique (CNRS), Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 (RID-AGE), 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), Charles University [Prague] (CU), AstraZeneca, European Project: 675179,H2020,H2020-MSCA-ITN-2015,TASPPI(2016), Landrieu, Isabelle, and Targeted small-molecule Stabilisation of Protein-Protein Interactions - TASPPI - - H20202016-02-01 - 2020-01-31 - 675179 - VALID

Subjects
0301 basic medicine, Magnetic Resonance Spectroscopy, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Protein Conformation, Dimer, Tau protein, tau Proteins, protein–protein interactions, Biochemistry, Epitope, Protein–protein interaction, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, NMR spectroscopy, Alzheimer Disease, Serine, Humans, Phosphorylation, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Molecular Biology, [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Binding Sites, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Chemistry, Drug discovery, 14-3-3 proteins, Cell Biology, Nuclear magnetic resonance spectroscopy, Surface Plasmon Resonance, Alzheimer's disease, Cyclic AMP-Dependent Protein Kinases, 030104 developmental biology, Monomer, Protein-Protein Interactions, 030220 oncology & carcinogenesis, Exoribonucleases, Mutation, Biophysics, biology.protein, Protein Multimerization, Alzheimer’s disease, analytical ultracentrifugation, Protein Binding
Abstract

Protein-protein interactions (PPIs) remain poorly explored targets for the treatment of Alzheimer’s disease (AD). The interaction of 14-3-3 proteins with Tau was shown to have detrimental effects on neuronal cells and to be linked to Tau pathology. This PPI is therefore seen as a potential target for AD. When Tau is phosphorylated by PKA (Tau-PKA), two 14-3-3 binding epitopes are generated, surrounding the phosphorylated serines 214 and 324 of Tau. The crystal structures of 14-3-3 in complex with peptides surrounding these Tau phosphosites show that both these motifs are anchored in the amphipathic binding groove of 14-3-3. However, in the absence of structural data with the full-length Tau protein, the stoichiometry of the complex or the interface and affinity of the partners, are still unclear. In this work, we addressed these points, using a broad range of biophysical techniques. The interaction of the long disordered Tau-PKA protein with 14-3-3σ is restricted to two short sequences, containing phosphorylated serines, which bind in the amphipathic binding groove of 14-3-3. Phosphorylation of Tau is fundamental for the formation of this stable complex, and the affinity of the Tau-PKA/14-3-3 interaction is in the 1-10 micromolar range. Each monomer of the 14-3-3σ dimer binds one of the two different phosphorylated peptides of Tau-PKA, suggesting a 14-3-3/Tau-PKA stoichiometry of 2:1, confirmed by analytical ultracentrifugation. These results contribute to a better understanding of this PPI and provide useful insights for drug discovery projects aiming at the inhibition of this interaction.

Published
2021

4. The O-β-linked N-acetylglucosaminylation of the Lamin B receptor and its impact on DNA binding and phosphorylation

Author

Caroline Smet-Nocca, Eleni Nikolakaki, Isabelle Landrieu, Adeline Page, François-Xavier Cantrelle, Thomas Giannakouros, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS), Aristotelian University of Thessaloniki, Aristotle University of Thessaloniki, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Turkeys, Glycosylation, Proto-Oncogene Proteins c-akt, [SDV]Life Sciences [q-bio], Biophysics, Receptors, Cytoplasmic and Nuclear, Lamin B receptor, N-Acetylglucosaminyltransferases, Biochemistry, Acetylglucosamine, 03 medical and health sciences, CDC2 Protein Kinase, Animals, Humans, Inner membrane, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Protein phosphorylation, Amino Acid Sequence, Phosphorylation, Nuclear protein, Molecular Biology, Integral membrane protein, ComputingMilieux_MISCELLANEOUS, Binding Sites, Sequence Homology, Amino Acid, Chemistry, DNA, Rats, Chromatin, Cell biology, 030104 developmental biology, Peptides, Protein Binding
Abstract

Lamin B Receptor (LBR) is an integral protein of the interphase inner nuclear membrane that is implicated in chromatin anchorage to the nuclear envelope. Phosphorylation of a stretch of arginine-serine (RS) dipeptides in the amino-terminal nucleoplasmic domain of LBR regulates the interactions of the receptor with other nuclear proteins, DNA and RNA and thus modulates tethering of heterochromatin to the nuclear envelope. While phosphorylation has been extensively studied, very little is known about other post-translational modifications of the protein. There is only one report on the O-β-linked N-acetyl-glucosaminylation (O-GlcNAcylation) of a serine residue downstream of the RS domain of rat LBR. In the present study we identify additional O-GlcNAcylation sites by using as substrates of O-β-N-acetylglucosaminyltransferase (OGT) a set of peptides containing the entire LBR RS domain or parts of it as well as flanking sequences. The in vitro activity of OGT was assessed by tandem mass spectrometry and NMR spectroscopy. Furthermore, we provide evidence that O-GlcNAcylation hampers DNA binding while it marginally affects RS domain phosphorylation mediated by SRPK1, Akt2 and cdk1 kinases. General significance Our methodology providing a quantitative description of O-GlcNAc patterns based on a combination of mass spectrometry and high resolution NMR spectroscopy on short peptide substrates allows subsequent functional analyses. Hence, our approach is of general interest to a wide audience of biologists aiming at deciphering the functional role of O-GlcNAc glycosylation and its crosstalk with phosphorylation.

Published
2018

5. Solution Structure of the N-Terminal Domain of Mediator Subunit MED26 and Molecular Characterization of Its Interaction with EAF1 and TAF7

Author

Zoé Lens, Xavier Hanoulle, Frédérique Dewitte, François-Xavier Cantrelle, Riccardo Peruzzini, Isabelle Landrieu, Marc Aumercier, Elisabeth Ferreira, Jean-Luc Baert, Vincent Villeret, Didier Monté, Alexis Verger, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), The NMR facilities were funded by the Hauts-de-France Regional Council, CNRS, Pasteur Institute of Lille, European Community (FEDER), French Research Ministry and the University of Sciences and Technologies of Lille I. We acknowledge support from the TGE RMN THC (FR-3050, France), FRABio (FR-3688, France), 'Projets Emergents' Nord-Pas-de-Calais Regional Council and Lille NMR and RPE Health and Biology core facility. R.P. was funded by a fellowship from Nord-Pas-de-Calais Regional Council., The FP7 WeNMR (project no. 261572) and H2020 West-Life (project no. 675858) European e-Infrastructure projects are acknowledged for the use of their Web portals., Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), ANR-16-CE12-0021,MEDNET,Le complexe Médiateur: d'un réseau d'interactions protéine-protéine à la régulation génique in vivo(2016), and Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Magnetic Resonance Spectroscopy, Protein Conformation, Protein subunit, RNA polymerase II, Biology, MESH: Transcription Factor TFIID/metabolism, 03 medical and health sciences, chemistry.chemical_compound, NMR spectroscopy, MESH: Protein Conformation, Structural Biology, Transcription (biology), RNA polymerase, Protein Interaction Mapping, Humans, MESH: Mediator Complex/metabolism, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, protein structure, Molecular Biology, TATA-Binding Protein Associated Factors, MESH: Transcription Factors/metabolism, MESH: Humans, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MEDiator complex, MESH: Magnetic Resonance Spectroscopy, MESH: Protein Interaction Mapping, MED26, MESH: Transcription Factor TFIID/chemistry, TAF7, Dissociation constant, Crystallography, MESH: Mediator Complex/chemistry, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], 030104 developmental biology, protein–protein interaction, chemistry, MESH: TATA-Binding Protein Associated Factors/chemistry, MESH: TATA-Binding Protein Associated Factors/metabolism, Helix, biology.protein, Biophysics, Transcription Factor TFIID, transcription regulation, Protein Binding, Transcription Factors
Abstract

International audience; MED26 is a subunit of Mediator, a large complex central to the regulation of gene transcription by RNA Polymerase II. MED26 plays a role in the switch between the initiation and elongation phases of RNA Polymerase II-mediated transcription process. Regulation of these steps requires successive binding of MED26 N-terminal domain (NTD) to TATA-binding protein-associated factor 7 (TAF7) and Eleven-nineteen lysine-rich in leukemia-Associated Factor 1 (EAF1). In order to investigate the mechanism of regulation by MED26, MED26-NTD structure was solved by NMR, revealing a 4-helix bundle. EAF1 (239-268) and TAF7 (205-235) peptide interactions were both mapped to the same groove formed by H3 and H4 helices of MED26-NTD. Both interactions are characterized by dissociation constants in the 10-μM range. Further experiments revealed a folding-upon-binding mechanism that leads to the formation of EAF1 (N247-S260) and TAF7 (L214-S227) helices. Chemical shift perturbations and nuclear Overhauser enhancement contacts support the involvement of residues I222/F223 in anchoring TAF7 helix to a hydrophobic pocket of MED26-NTD, including residues L48, W80 and I84. In addition, Ala mutations of charged residues located in the C-terminal disordered part of TAF7 and EAF1 peptides affected the binding, with a loss of affinity characterized by a 10-time increase of dissociation constants. A structural model of MED26-NTD/TAF7 complex shows bi-partite components, combining ordered and disordered segments, as well as hydrophobic and electrostatic contributions to the binding. This study provides molecular detail that will help to decipher the mechanistic basis for the initiation to elongation switch-function mediated by MED26-NTD.

Published
2017

6. Regulation of the interaction between the neuronal BIN1 isoform 1 and Tau proteins - role of the SH3 domain

Author

Isabelle Landrieu, François-Xavier Cantrelle, Yoann Sottejeau, Guy Lippens, Idir Malki, Jean-Charles Lambert, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP), The NMR facilities were funded by the Nord Region Council, CNRS, Pasteur Institute of Lille, European Community (FEDER), French Research Ministry and Lille University. We acknowledge support from TGE RMN THC (FR-3050, France) and FRABio (Univ. Lille, CNRS, FR 3688). This study was supported by a grant from the LabEx (Laboratory of Excellence), DISTALZ (Development of Innovative Strategies for a Transdisciplinary approach to Alzheimer's disease), by the French government funding agency Agence Nationale de la Recherche ANR-15-CE16-0002 BinAlz and by the Alzheimer's association (BFG-14-318355)., Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), CNRS, Université de Lille, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 [UGSF], Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 [RID-AGE], and Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]

Subjects
0301 basic medicine, Models, Molecular, Protein Conformation, alpha-Helical, Alzheimer’s disease, BIN1, nuclearmagnetic resonance spectroscopy, protein–protein interaction, SH3 domain, Tau, MESH: Sequence Homology, Amino Acid, MESH: Adaptor Proteins, Signal Transducing/metabolism, Amino Acid Motifs, MESH: Tumor Suppressor Proteins/genetics, Gene Expression, Peptide, Biochemistry, MESH: Amino Acid Motifs, 0302 clinical medicine, MESH: Adaptor Proteins, Signal Transducing/genetics, MESH: Nuclear Magnetic Resonance, Biomolecular, Protein Isoforms, Cloning, Molecular, MESH: tau Proteins/chemistry, chemistry.chemical_classification, Neurons, biology, MESH: Kinetics, Chemistry, Nuclear Proteins, MESH: Neurons/metabolism, Nuclear magnetic resonance spectroscopy, MESH: Peptides/genetics, MESH: Tumor Suppressor Proteins/chemistry, MESH: tau Proteins/metabolism, Recombinant Proteins, MESH: Tumor Suppressor Proteins/metabolism, MESH: Recombinant Proteins/chemistry, MESH: tau Proteins/genetics, MESH: Peptides/metabolism, Domain (ring theory), MESH: Protein Conformation, beta-Strand, MESH: Protein Isoforms/metabolism, MESH: Models, Molecular, MESH: Nuclear Proteins/genetics, Protein Binding, Gene isoform, MESH: Recombinant Proteins/metabolism, MESH: Gene Expression, MESH: Sequence Alignment, tau Proteins, MESH: Escherichia coli/genetics, Clathrin, Protein–protein interaction, MESH: Peptides/chemistry, 03 medical and health sciences, Escherichia coli, Humans, MESH: Protein Binding, Protein Interaction Domains and Motifs, MESH: Adaptor Proteins, Signal Transducing/chemistry, MESH: Recombinant Proteins/genetics, MESH: Cloning, Molecular, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Nuclear Proteins/metabolism, Molecular Biology, Gene, Nuclear Magnetic Resonance, Biomolecular, Adaptor Proteins, Signal Transducing, MESH: Protein Conformation, alpha-Helical, MESH: Protein Interaction Domains and Motifs, Binding Sites, MESH: Protein Isoforms/genetics, MESH: Humans, Sequence Homology, Amino Acid, Tumor Suppressor Proteins, Cell Biology, MESH: Nuclear Proteins/chemistry, Kinetics, MESH: Neurons/chemistry, 030104 developmental biology, MESH: Binding Sites, MESH: Protein Isoforms/chemistry, Biophysics, biology.protein, MESH: Escherichia coli/metabolism, Protein Conformation, beta-Strand, Peptides, Sequence Alignment, 030217 neurology & neurosurgery
Abstract

International audience; Bridging integrator 1 (bin1) gene is a genetic determinant of Alzheimer's disease (AD) and has been reported to modulate Alzheimer's pathogenesis through pathway(s) involving Tau. The functional impact of Tau/BIN1 interaction as well as the molecular details of this interaction are still not fully resolved. As a consequence, how BIN1 through its interaction with Tau affects AD risk is also still not determined. To progress in this understanding, interaction of Tau with two BIN1 isoforms was investigated using Nuclear Magnetic Resonance spectroscopy. 1H, 15N spectra showed that the C-terminal SH3 domain of BIN1 isoform 1 (BIN1Iso1) is not mobile in solution but locked with the core of the protein. In contrast, the SH3 domain of BIN1 isoform 9 (BIN1Iso9) behaves as an independent mobile domain. This reveals an equilibrium between close and open conformations for the SH3 domain. Interestingly, a 334–376 peptide from the clathrin and AP-2-binding domain (CLAP) domain of BIN1Iso1, which contains a SH3-binding site, is able to compete with BIN1-SH3 intramolecular interaction. For both BIN1 isoforms, the SH3 domain can interact with Tau(210–240) sequence. Tau(210–240) peptide can indeed displace the intramolecular interaction of the BIN1-SH3 of BIN1Iso1 and form a complex with the released domain. The measured Kd were in agreement with a stronger affinity of Tau peptide. Both CLAP and Tau peptides occupied the same surface on the BIN1-SH3 domain, showing that their interaction is mutually exclusive. These results emphasize an additional level of complexity in the regulation of the interaction between BIN1 and Tau dependent of the BIN1 isoforms.

Published
2017

7. Identification of the Tau phosphorylation pattern that drives its aggregation

Author

François-Xavier Cantrelle, Haoling Qi, Yves Jacquot, Guy Lippens, Isabelle Huvent, Isabelle Landrieu, Cillian Byrne, Clément Despres, Béatrice Chambraud, Etienne-Emile Baulieu, Caroline Smet-Nocca, CNRS, Université de Lille, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF], Laboratoire des biomolécules [LBM UMR 7203], Université Pierre et Marie Curie - Paris 6 [UPMC], Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 [UGSF], Petites Molécules de neuroprotection, neurorégénération et remyélinisation, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés [LISBP], Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC), Neuroprotection et neurogénération (UMR788), Neuroprotection et Neurodégénération (UMR788), TGE RMN THC (Tres Grands Equipements Resonance Magnetique Nucleaire Tres Hauts Champs, France) [FR-3050], FRABio (Federation de Recherche 'Biochimie Structurale & Fonctionnelle des Assemblages Biomoleculaires', France [FR 3688], MetaToul [MetaboHUB-AR-11-INBS-0010], Université de Lille-Centre National de la Recherche Scientifique (CNRS), Landrieu, Isabelle, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), and Lippens, Guy

Subjects
0301 basic medicine, Models, Molecular, Threonine, inorganic chemicals, Phosphorylation sites, Magnetic Resonance Spectroscopy, Tau, phosphorylation, aggregation, Alzheimer's disease, NMR, tau Proteins, Biotechnologies, macromolecular substances, Protein Aggregation, Pathological, environment and public health, Rats, Sprague-Dawley, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Microscopy, Electron, Transmission, Protein Domains, Alzheimer’s disease, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Serine, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, ComputingMilieux_MISCELLANEOUS, Multidisciplinary, Binding Sites, Kinase, Nuclear magnetic resonance spectroscopy, Biological Sciences, Fluorescence, In vitro, Peptide Fragments, 3. Good health, enzymes and coenzymes (carbohydrates), 030104 developmental biology, chemistry, Biochemistry, Tau phosphorylation, Phosphorylation, bacteria, Thioflavin, 030217 neurology & neurosurgery
Abstract

International audience; Determining the functional relationship between Tau phosphorylation and aggregation has proven a challenge owing to the multiple potential phosphorylation sites and their clustering in the Tau sequence. We use here in vitro kinase assays combined with NMR spectroscopy as an analytical tool to generate well-characterized phosphorylated Tau samples and show that the combined phosphorylation at the Ser202/Thr205/Ser208 sites, together with absence of phosphorylation at the Ser262 site, yields a Tau sample that readily forms fibers, as observed by thioflavin T fluorescence and electron microscopy. On the basis of conformational analysis of synthetic phosphorylated peptides, we show that aggregation of the samples correlates with destabilization of the turn-like structure defined by phosphorylation of Ser202/Thr205.

Published
2017

8. A functional fragment of Tau forms fibers without the need for an intermolecular cysteine bridge

Author

Amina Kamah, Isabelle Landrieu, Isabelle Huvent, Guy Lippens, Christian Slomianny, François-Xavier Cantrelle, Nicolas Barois, Caroline Smet-Nocca, Université Lille Nord de France (COMUE), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Plateforme BioImaging Center Lille (BICeL), Plateformes Lilloises en Biologie et Santé - UMS 2014 - US 41 (PLBS), 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)-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), Laboratoire de Physiologie Cellulaire : Canaux ioniques, inflammation et cancer - U 1003 (PHYCELL), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université de Lille, Plate-forme BICeL (IFR142), Physiologie Cellulaire (PHYCEL) - U1003 (PHYCEL), Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE), Université de Lille-Centre National de la Recherche Scientifique (CNRS), and Plateformes Lilloises en Biologie et Santé - UAR 2014 - US 41 (PLBS)

Subjects
Magnetic Resonance Spectroscopy, Biophysics, tau Proteins, macromolecular substances, Fibril, Biochemistry, law.invention, Aggregation, 03 medical and health sciences, chemistry.chemical_compound, NMR spectroscopy, 0302 clinical medicine, law, Microtubule, mental disorders, Electron microscopy, Humans, Cysteine, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Binding site, Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Heparin, Cell Biology, Nuclear magnetic resonance spectroscopy, Fluorescence, Recombinant Proteins, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Crystallography, nervous system, chemistry, Nucleation, Thioflavin, Tau, Electron microscope, 030217 neurology & neurosurgery
Abstract

International audience; We study the aggregation of a fragment of the neuronal protein Tau that contains part of the proline rich domain and of the microtubule binding repeats. When incubated at 37 °C with heparin, the fragment readily forms fibers as witnessed by Thioflavin T fluorescence. Electron microscopy and NMR spectroscopy show bundled ribbon like structures with most residues rigidly incorporated in the fibril. Without its cysteines, this fragment still forms fibers of a similar morphology, but with lesser Thioflavin T binding sites and more mobility for the C-terminal residues.

Published
2014

9. Investigation of Homeodomain Membrane Translocation Properties: Insights from the Structure Determination of Engrailed-2 Homeodomain in Aqueous and Membrane-Mimetic Environments

Author

Olivier Lequin, Alain Joliot, Lucie Khemtémourian, Stéphane Balayssac, Ludovic Carlier, François-Xavier Cantrelle, Gérard Chassaing, Université Pierre et Marie Curie - Paris 6 (UPMC), Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique (SPCMIB), 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-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie des Substances Naturelles (ICSN), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), 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), Synthèse, Structure et Fonction de Molécules Bioactives (SSFMB), Université Pierre et Marie Curie - Paris 6 (UPMC)-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), and Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Conformational change, Circular dichroism, media_common.quotation_subject, Molecular Sequence Data, Biophysics, Nerve Tissue Proteins, Biology, 010402 general chemistry, 01 natural sciences, Micelle, 03 medical and health sciences, Animals, Humans, Amino Acid Sequence, Internalization, Micelles, 030304 developmental biology, media_common, Homeodomain Proteins, 0303 health sciences, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Cell Membrane, Membrane, Biological membrane, engrailed, 0104 chemical sciences, Protein Structure, Tertiary, Crystallography, Protein Transport, Helix, Drosophila, Chickens
Abstract

International audience; In addition to their well-known DNA-binding properties, homeodomains have the ability to efficiently translocate across biological membranes through still poorly-characterized mechanisms. To date, most biophysical studies addressing the mechanisms of internalization have focused on small synthetic peptides rather than full-length globular homeodomains. In this work, we characterized the conformational properties of chicken Engrailed 2 homeodomain (En2HD) in aqueous solution and in membrane mimetic environments using circular dichroism, Trp fluorescence, and NMR spectroscopy. En2HD adopts a well-defined three-helical bundle fold in aqueous solution. The Trp-48 residue, which is critical for internalization, is fully buried in the hydrophobic core. Circular dichroism and fluorescence reveal that a conformational transition occurs in anionic lipid vesicles and in micelles. En2HD loses its native three-dimensional structure in micellar environments but, remarkably, near-native helical secondary structures are maintained. Long-range interactions could be detected using site-directed spin labels, indicating that the three helices do not adopt extended orientations. Noncovalent paramagnetic probes yielded information about helix positioning and unveiled the burial of critical aromatic and basic residues within the micelles. Our results suggest that electrostatic interactions with membranes may be determinant in inducing a conformational change enabling Trp-48 to insert into membranes.

Published
2013
Full Text
View/download PDF

10. Characterization of Neuronal Tau Protein as a Target of Extracellular Signal-regulated Kinase

Author

Haoling Qi, Jeremy Gunawardena, Guy Lippens, Béatrice Chambraud, François-Xavier Cantrelle, Sudhakaran Prabakaran, Isabelle Landrieu, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Department of Systems Biology, Harvard Medical School [Boston] (HMS), Neuroprotection et neurogénération (UMR788), Neuroprotection et Neurodégénération (UMR788), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), CNRS, Université de Lille, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF], Harvard Medical School [Boston] [HMS], Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 [UGSF], Petites Molécules de neuroprotection, neurorégénération et remyélinisation, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Landrieu, Isabelle, Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Prabakaran, Sudhakaran [0000-0002-6527-1085], and Apollo - University of Cambridge Repository

Subjects
0301 basic medicine, MAPK/ERK pathway, Amino Acid Motifs, spectroscopie rmn, Biochemistry, environment and public health, protein-protein interaction, neurodégénérescence, Protein phosphorylation, Phosphorylation, Extracellular Signal-Regulated MAP Kinases, [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Kinase, Neurodegeneration, Molecular Bases of Disease, 3. Good health, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], maladie d'alzheimer, extracellular signal-regulated kinase (ERK), protéine, cerveau, Tau protein (Tau), Alzheimer's disease, Alzheimer disease, inorganic chemicals, nuclear magnetic resonance (NMR), Tau protein, Médecine humaine et pathologie, Hyperphosphorylation, tau Proteins, Biology, 03 medical and health sciences, interaction Tau protein (Tau), mental disorders, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, protein phosphorylation, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], protein phosphorylation protein-protein, Nuclear Magnetic Resonance, Biomolecular, Molecular Biology, Cell Biology, medicine.disease, Protein Structure, Tertiary, Rats, enzymes and coenzymes (carbohydrates), 030104 developmental biology, biology.protein, Human health and pathology
Abstract

International audience; Tau neuronal protein has a central role in neurodegeneration and is implicated in Alzheimer disease development. Abnormal phosphorylation of Tau impairs its interaction with other proteins and is associated with its dysregulation in pathological conditions. Molecular mechanisms leading to hyperphosphorylation of Tau in pathological conditions are unknown. Here, we characterize phosphorylation of Tau by extracellular-regulated kinase (ERK2), a mitogen-activated kinase (MAPK) that responds to extracellular signals. Analysis of in vitro phosphorylated Tau by activated recombinant ERK2 with nuclear magnetic resonance spectroscopy (NMR) reveals phosphorylation of 15 Ser/Thr sites. In vitro phosphorylation of Tau using rat brain extract and subsequent NMR analysis identifies the same sites. Phosphorylation with rat brain extract is known to transform Tau into an Alzheimer disease-like state. Our results indicate that phosphorylation of Tau by ERK2 alone is sufficient to produce the same characteristics. We further investigate the mechanism of ERK2 phosphorylation of Tau. Kinases are known to recognize their protein substrates not only by their specificity for a targeted Ser or Thr phosphorylation site but also by binding to linear-peptide motifs called docking sites. We identify two main ERK2 docking sites in Tau sequence using NMR. Our results suggest that ERK2 dysregulation in Alzheimer disease could lead to abnormal phosphorylation of Tau resulting in the pathology of the disease.

Published
2016

11. Structural basis for oxygen degradation domain selectivity of the HIF prolyl hydroxylases

Author

Rasheduzzaman, Chowdhury, Ivanhoe K H, Leung, Ya-Min, Tian, Martine I, Abboud, Wei, Ge, Carmen, Domene, François-Xavier, Cantrelle, Isabelle, Landrieu, Adam P, Hardy, Christopher W, Pugh, Peter J, Ratcliffe, Timothy D W, Claridge, and Christopher J, Schofield

Subjects
Proline, Polycythemia, Fibroblasts, Molecular Dynamics Simulation, Crystallography, X-Ray, Hydroxylation, Hypoxia-Inducible Factor 1, alpha Subunit, Recombinant Proteins, Article, Hypoxia-Inducible Factor-Proline Dioxygenases, Isoenzymes, Oxygen, Mice, Structure-Activity Relationship, Protein Domains, Neoplasms, Animals, Humans, Hypoxia, Cells, Cultured, Protein Binding
Abstract

The response to hypoxia in animals involves the expression of multiple genes regulated by the αβ-hypoxia-inducible transcription factors (HIFs). The hypoxia-sensing mechanism involves oxygen limited hydroxylation of prolyl residues in the N- and C-terminal oxygen-dependent degradation domains (NODD and CODD) of HIFα isoforms, as catalysed by prolyl hydroxylases (PHD 1–3). Prolyl hydroxylation promotes binding of HIFα to the von Hippel–Lindau protein (VHL)–elongin B/C complex, thus signalling for proteosomal degradation of HIFα. We reveal that certain PHD2 variants linked to familial erythrocytosis and cancer are highly selective for CODD or NODD. Crystalline and solution state studies coupled to kinetic and cellular analyses reveal how wild-type and variant PHDs achieve ODD selectivity via different dynamic interactions involving loop and C-terminal regions. The results inform on how HIF target gene selectivity is achieved and will be of use in developing selective PHD inhibitors., The response to hypoxia involves multiple genes regulated by the hypoxia inducible transcription factors (HIFs), whose stability is regulated by prolyl hydroxylation. Here the authors provide a molecular basis for the substrate selectivity of the HIF prolyl hydroxylases that can be altered in erythrocytosis and cancer.

Published
2016

12. Isomerisation and oligomerization of truncated and mutated tau forms by FKBP52 are independent processes

Author

Julien Giustiniani, Yves Jacquot, Isabelle Landrieu, Béatrice Chambraud, Guy Lippens, François-Xavier Cantrelle, Caroline Smet, Etienne-Emile Baulieu, Isabelle Huvent, Kevin Guillemeau, Amina Kamah, Cillian Byrne, Université Lille Nord de France (COMUE), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut de Recherche Interdisciplinaire [Villeneuve d'Ascq] (IRI), Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Droit et Santé-Université de Lille, Sciences et Technologies, Neuroprotection et neurogénération (UMR788), Neuroprotection et Neurodégénération (UMR788), Peptides, glycoconjugués et métaux en biologie (LBM-E1), Laboratoire des biomolécules (LBM UMR 7203), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC), ANR (program MALZ-TAF), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université de Lille, Sciences et Technologies-Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Lille-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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
0301 basic medicine, Isomerase activity, Tau protein, Mutant, tau Proteins, Peptide, Protein Aggregation, Pathological, Tacrolimus Binding Proteins, 03 medical and health sciences, 0302 clinical medicine, Isomerism, Structural Biology, mental disorders, Humans, Molecular Biology, FK506 binding, chemistry.chemical_classification, biology, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, aggregation, Alzheimer's disease, FKBP52, prolyl cis/trans isomerase activity, 030104 developmental biology, FKBP, Biochemistry, Biophysics, biology.protein, Protein Multimerization, Tau, 030217 neurology & neurosurgery, Heteronuclear single quantum coherence spectroscopy
Abstract

International audience; The aggregation of the neuronal Tau protein is one molecular hallmark of Alzheimer's disease and other related tauopathies, but the precise molecular mechanisms of the aggregation process remain unclear. The FK506 binding protein FKBP52 is able to induce oligomers in the pathogenic Tau P301L mutant and in a truncated form of the wild-type human Tau protein. Here, we investigate whether FKBP52's capacity to induce Tau oligomers depends on its prolyl cis/trans isomerase activity. We find that FKBP52 indeed can isomerize selected prolyl bonds in the different Tau proteins, and that this activity is carried solely by its first FK506 binding domain. Its capacity to oligomerize Tau is however not linked to this PPIase activity. In addition, we identified a novel molecular interaction implying the PHF6 peptide of Tau and the FK1/FK2 domains of FKBP52 independent of FK506 binding; these data point towards a non-catalytic molecular interaction that might govern the effect of FKBP52 on Tau.

Published
2016

13. Proline Conformation in a Functional Tau Fragment

Author

Puneet Ahuja, Guy Lippens, Caroline Smet, Jean-Michel Wieruszeski, Juan José Marín López, Xavier Hanoulle, Isabelle Landrieu, François-Xavier Cantrelle, Isabelle Huvent, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université de Lille, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), and Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Magnetic Resonance Spectroscopy, Proline, Stereochemistry, Protein Conformation, Tau protein, tau Proteins, Isomerase, 03 medical and health sciences, Protein structure, Structural Biology, Humans, Phosphorylation, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Molecular Biology, Conformational isomerism, ComputingMilieux_MISCELLANEOUS, biology, Chemistry, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Folding (chemistry), 030104 developmental biology, biology.protein, Two-dimensional nuclear magnetic resonance spectroscopy, Protein Processing, Post-Translational, Heteronuclear single quantum coherence spectroscopy
Abstract

The conformational state of distinct prolines can determine the folding of a protein but equally other biological processes when coupled to a conformation-sensitive secondary reaction. For the neuronal tau protein, the importance of proline conformation is underscored by its interaction with different prolyl cis/trans isomerases. The proline conformation would gain even further importance after phosphorylation of the preceding residue by various proline-directed kinases. A number of molecular diseases including Alzheimer's disease and traumatic brain injury were thereby recently qualified as "cistauosis", as they would imply a cis conformation for the pThr231-Pro232 prolyl bond. We here investigate by NMR spectroscopy the conformation of all prolines in a functional Tau fragment, Tau[208-324]. Although we can detect and identify some minor conformers in the cis form, we show that all prolines are for over 90% in the trans conformation. Phosphorylation by CDK2/CycA3, which notably leads to complete modification of the Thr231 residue, does not change this conclusion. Our data hence disagree with the notion that specific prolyl bonds in tau would adopt preferentially the cis conformation.

Published
2016

14. Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

Author

Haoling Qi, Robert Schneider, Xavier Hanoulle, Hamida Merzougui, Isabelle Landrieu, François-Xavier Cantrelle, Clément Despres, Clément Danis, Caroline Smet-Nocca, Luiza M. Bessa, Idir Malki, Guy Lippens, Isabelle Huvent, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer (JPArc - U837 Inserm), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université Lille 2 - Faculté de Médecine, CNRS, Université de Lille, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 [UGSF], Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U837 [JPArc], Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF], Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U1172 Inserm - U837 (JPArc), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE)-Université de Lille, Université de Lille-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U837 (JPArc), and Université Lille Nord de France (COMUE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille

Subjects
0301 basic medicine, Magnetic Resonance Spectroscopy, General Chemical Engineering, Tau protein, tau Proteins, Intrinsically disordered proteins, Biochemistry, General Biochemistry, Genetics and Molecular Biology, Isotopic labeling, 03 medical and health sciences, Alzheimer Disease, Protein purification, mental disorders, Triple-resonance nuclear magnetic resonance spectroscopy, Humans, Protein phosphorylation, Phosphorylation, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], ComputingMilieux_MISCELLANEOUS, biology, General Immunology and Microbiology, Chemistry, General Neuroscience, Nuclear magnetic resonance spectroscopy, Recombinant Proteins, 3. Good health, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Intrinsically Disordered Proteins, 030104 developmental biology, biology.protein, Issue 118, nuclear magnetic resonance spectroscopy, isotopic labeling, intrinsically disordered protein, recombinantprotein, protein purification, protein phosphorylation, NMR data acquisition, resonance assignment
Abstract

Aggregates of the neuronal Tau protein are found inside neurons of Alzheimer's disease patients. Development of the disease is accompanied by increased, abnormal phosphorylation of Tau. In the course of the molecular investigation of Tau functions and dysfunctions in the disease, nuclear magnetic resonance (NMR) spectroscopy is used to identify the multiple phosphorylations of Tau. We present here detailed protocols of recombinant production of Tau in bacteria, with isotopic enrichment for NMR studies. Purification steps that take advantage of Tau's heat stability and high isoelectric point are described. The protocol for in vitro phosphorylation of Tau by recombinant activated ERK2 allows for generating multiple phosphorylations. The protein sample is ready for data acquisition at the issue of these steps. The parameter setup to start recording on the spectrometer is considered next. Finally, the strategy to identify phosphorylation sites of modified Tau, based on NMR data, is explained. The benefit of this methodology compared to other techniques used to identify phosphorylation sites, such as immuno-detection or mass spectrometry (MS), is discussed. 118

Published
2016

15. Tau phosphorylation regulates the interaction between BIN1's SH3 domain and Tau's proline-rich domain

Author

Yoann, Sottejeau, Alexis, Bretteville, François-Xavier, Cantrelle, Nicolas, Malmanche, Florie, Demiaute, Tiago, Mendes, Charlotte, Delay, Harmony, Alves Dos Alves, Amandine, Flaig, Peter, Davies, Pierre, Dourlen, Bart, Dermaut, Jocelyn, Laporte, Philippe, Amouyel, Guy, Lippens, Julien, Chapuis, Isabelle, Landrieu, and Jean-Charles, Lambert

Subjects
Neurons, Magnetic Resonance Spectroscopy, Proline, Protein Conformation, Tumor Suppressor Proteins, Research, Brain, Nuclear Proteins, tau Proteins, Transfection, Rats, src Homology Domains, HEK293 Cells, Animals, Newborn, Animals, Humans, Phosphorylation, Cells, Cultured, Adaptor Proteins, Signal Transducing
Abstract

Introduction The application of high-throughput genomic approaches has revealed 24 novel risk loci for Alzheimer’s disease (AD). We recently reported that the bridging integrator 1 (BIN1) risk gene is linked to Tau pathology. Results We used glutathione S-transferase pull-down assays and nuclear magnetic resonance (NMR) experiments to demonstrate that BIN1 and Tau proteins interact directly and then map the interaction between BIN1’s SH3 domain and Tau’s proline-rich domain (PRD) . Our NMR data showed that Tau phosphorylation at Thr231 weakens the SH3-PRD interaction. Using primary neurons, we found that BIN1-Tau complexes partly co-localize with the actin cytoskeleton; however, these complexes were not observed with Thr231-phosphorylated Tau species. Conclusion Our results show that (i) BIN1 and Tau bind through an SH3-PRD interaction and (ii) the interaction is downregulated by phosphorylation of Tau Thr231 (and potentially other residues). Our study sheds new light on regulation of the BIN1/Tau interaction and opens up new avenues for exploring its complex’s role in the pathogenesis of AD. Electronic supplementary material The online version of this article (doi:10.1186/s40478-015-0237-8) contains supplementary material, which is available to authorized users.

Published
2015

16. A Phosphorylation-Induced Turn Defines the Alzheimer's Disease AT8 Antibody Epitope on the Tau Protein

Author

Predrag Kukic, Yves Jacquot, Isabelle Landrieu, Guy Lippens, Neha S. Gandhi, Ricardo L. Mancera, Jean-Michel Wieruszeski, Laziza Amniai, Cillian Byrne, François-Xavier Cantrelle, Curtin University [Perth], Planning and Transport Research Centre (PATREC), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC), Institut Baulieu, Department of Chemistry [Cambridge, UK], University of Cambridge [UK] (CAM), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), CNRS, Université de Lille, Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576, Laboratoire des biomolécules [LBM UMR 7203], University of Cambridge [UK] [CAM], and Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 [UGSF]

Subjects
Tau protein, tau Proteins, Molecular Dynamics Simulation, Catalysis, Epitope, Turn (biochemistry), Epitopes, 03 medical and health sciences, 0302 clinical medicine, Alzheimer Disease, Humans, Phosphorylation, Nuclear Magnetic Resonance, Biomolecular, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Linear epitope, biology, Chemistry, Alzheimer’s disease, AT8 antibody, epitope mapping, phosphorylation, tau protein, Antibodies, Monoclonal, General Chemistry, Nuclear magnetic resonance spectroscopy, General Medicine, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, Epitope mapping, Biochemistry, biology.protein, Antibody, Epitope Mapping, 030217 neurology & neurosurgery
Abstract

Post mortem biochemical staging of Alzheimer’s disease is currently based on immunochemical analysis of brain slices with the AT8 antibody. The epitope of AT8 is described around the pSer202/pThr205 region of the hyperphosphorylated form of the neuronal protein tau. In this study, NMR spectroscopy was used to precisely map the AT8 epitope on phosphorylated tau, and derive its defining structural features by a combination of NMR analyses and molecular dynamics. A particular turn conformation is stabilized by a hydrogen bond of the phosphorylated Thr205 residue to the amide proton of Gly207, and is further stabilized by the two Arg residues opposing the pSer202/pThr205. 54;23

Published
2015

17. Identification of a Plasmodium falciparum inhibitor-2 motif involved in the binding and regulation activity of protein phosphatase type 1

Author

François-Xavier Cantrelle, Jamal Khalife, Isabelle Landrieu, Alain Martoriati, Katia Cailliau-Maggio, Jérôme Vicogne, Aline Fréville, Géraldine Tellier, Christine Pierrot, Audrey Vandomme, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), 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, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Institut de biologie de Lille - IBL (IBLI), Université de Lille, Sciences et Technologies-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Droit et Santé, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université Lille Nord de France (COMUE), Laboratoire de Régulation des Signaux de Division, Université de Lille, Sciences et Technologies, The NMR facilities were funded by the Région Nord, CNRS, Pasteur Institute of Lille, European Community (FEDER), French Research Ministry and the University of Sciences and Technologies of Lille I. We gratefully acknowledge support from the TGIR‐RMN‐THC FR3050 CNRS., We thank Hadidjatou Kalamou for technical assistance. We thank Dr RJ Pierce for critical reading of the manuscript., Centre d’Infection et d’Immunité de Lille (CIIL) - U1019 - 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 National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), 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), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique (CNRS), and Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
Erythrocytes, Amino Acid Motifs, Drug Evaluation, Preclinical, Protozoan Proteins, Xenopus, Regulator, protein phosphatase type 1, Peptide, MESH: Amino Acid Sequence, MESH: Protein Phosphatase 1/physiology, Biochemistry, Homology (biology), MESH: Protozoan Proteins/chemistry, protein-protein interaction, Xenopus laevis, MESH: Amino Acid Motifs, Protein Phosphatase 1, MESH: Antimalarials/pharmacology, MESH: Animals, Enzyme Inhibitors, Cells, Cultured, MESH: Antimalarials/chemistry, chemistry.chemical_classification, 0303 health sciences, 030302 biochemistry & molecular biology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], MESH: Drug Evaluation, Preclinical, MESH: Plasmodium falciparum/enzymology, Protein Binding, MESH: Cells, Cultured, PP1 inhibitor 2, MESH: Plasmodium falciparum/drug effects, MESH: Protein Phosphatase 1/chemistry, Immunoprecipitation, Molecular Sequence Data, Phosphatase, Plasmodium falciparum, Biology, MESH: Protein Phosphatase 1/antagonists & inhibitors, Protein–protein interaction, Antimalarials, 03 medical and health sciences, NMR spectroscopy, MESH: Xenopus laevis, Animals, Humans, MESH: Protein Binding, Protein Interaction Domains and Motifs, Amino Acid Sequence, MESH: Enzyme Inhibitors/chemistry, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Molecular Biology, 030304 developmental biology, MESH: Protozoan Proteins/antagonists & inhibitors, MESH: Protein Interaction Domains and Motifs, Germinal vesicle, MESH: Humans, MESH: Molecular Sequence Data, MESH: Enzyme Inhibitors/pharmacology, MESH: Enzyme Inhibitors/metabolism, Cell Biology, biology.organism_classification, MESH: Antimalarials/metabolism, chemistry, MESH: Protozoan Proteins/physiology, MESH: Erythrocytes/parasitology
Abstract

The regulation of Plasmodium falciparum protein phosphatase type 1 (PfPP1) activity remains to be deciphered. Data from homologous eukaryotic type 1 protein phosphatases (PP1) suggest that several protein regulators should be involved in this essential process. One such regulator, named PfI2 based on its primary sequence homology with eukaryotic inhibitor 2 (I2), was recently shown to be able to interact with PfPP1 and to inhibit its phosphatase activity, mainly through the canonical ‘RVxF’ binding motif. The details of the structural and functional characteristics of this interaction are investigated here. Using NMR spectroscopy, a second site of interaction is suggested to reside between residues D94 and T117 and contains the ‘FxxR/KxR/K’ binding motif present in other I2 proteins. This site seems to play in concert/synergy with the ‘RVxF’ motif to bind PP1, because only mutations in both motifs were able to abolish this interaction completely. However, regarding the structure/function relationship, mutation of either the ‘RVxF’ or ‘FxxR/KxR/K’ motif is more drastic, because each mutation prevents the capacity of PfI2 to trigger germinal vesicle breakdown in microinjected Xenopus oocytes. This indicates that the tight association of the PfI2 regulator to PP1, mediated by a two-site interaction, is necessary to exert its function. Based on these results, the use of a peptide derived from the ‘FxxR/KxR/K’ PfI2 motif was investigated for its potential effect on Plasmodium growth. This peptide, fused at its N-terminus to a penetrating sequence, was shown to accumulate specifically in infected erythrocytes and to have an antiplasmodial effect. Structured digital abstract PfPP1 and PfI2 bind by nuclear magnetic resonance (1, 2) PP1 physically interacts with PfPP1 by anti tag coimmunoprecipitation (View interaction) PfI2 binds to PfPP1 by enzyme linked immunosorbent assay (View interaction) PfI2 binds to PfPP1 by surface plasmon resonance (1, 2, 3, 4)

Published
2014

18. Nuclear Magnetic Resonance Analysis of the Acetylation Pattern of the Neuronal Tau Protein

Author

Isabelle Landrieu, François-Xavier Cantrelle, Guy Lippens, Amina Kamah, Isabelle Huvent, Caroline Smet-Nocca, Haoling Qi, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université de Lille, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université Lille Nord de France (COMUE), and Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
Lysine, Tau protein, tau Proteins, Biochemistry, 03 medical and health sciences, 0302 clinical medicine, Nuclear magnetic resonance, Microtubule, Humans, Post-translational regulation, p300-CBP Transcription Factors, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Cysteine, Nuclear Magnetic Resonance, Biomolecular, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, biology, Nitrogen Isotopes, Chemistry, Heparin, Acetylation, Nuclear magnetic resonance spectroscopy, In vitro, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Kinetics, Acetyltransferase, biology.protein, Protein Processing, Post-Translational, 030217 neurology & neurosurgery
Abstract

International audience; Lysine acetylation of the neuronal Tau protein was described as a novel mechanism of posttranslational regulation of Tau functions with important outcomes in microtubule binding and aggregation processes related to Alzheimer's disease. Here, we unravel at a per-residue resolution the acetylation pattern of full-length Tau by the Creb-binding protein (CBP) acetyltransferase using high-resolution nuclear magnetic resonance spectroscopy. Our study gives a quantitative overview of CBP-mediated acetylation and examines the catalytic proficiency because the nonenzymatic reaction with acetyl-coenzyme A occurs in vitro. Furthermore, we have investigated with this characterized acetylated Tau the effect of acetylation on Tau fibrillization in a heparin-induced aggregation assay and on heparin binding.

Published
2014

19. Structure, function, and evolution of the beta-thymosin/WH2 (WASP-Homology2) actin-binding module

Author

Maud Hertzog, Carine van Heijenoort, Louis Renault, Marcel Knossow, Dominique Didry, Eric Guittet, Marie-France Carlier, François-Xavier Cantrelle, Institut de Chimie des Substances Naturelles (ICSN), and Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)

Subjects
Models, Molecular, MESH: Thymosin, Protein Conformation, Molecular Sequence Data, Plasma protein binding, macromolecular substances, MESH: Amino Acid Sequence, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH: Actins, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Protein structure, MESH: Protein Conformation, History and Philosophy of Science, Animals, Humans, MESH: Protein Binding, MESH: Animals, Actin-binding protein, Amino Acid Sequence, Peptide sequence, Actin, 030304 developmental biology, 0303 health sciences, Binding Sites, MESH: Humans, MESH: Molecular Sequence Data, biology, General Neuroscience, 030302 biochemistry & molecular biology, Actin remodeling, Actins, Cell biology, Thymosin, Profilin, MESH: Binding Sites, biology.protein, MDia1, Endothelium, Vascular, MESH: Endothelium, Vascular, Epidermis, MESH: Epidermis, MESH: Models, Molecular, Protein Binding
Abstract

International audience; beta-thymosins are acknowledged G-actin sequesterers. However, in the recent years, the conserved beta-thymosins/WH2 actin-binding module, has been identified in a large number of proteins that all interact with actin and play diverse functions in cell motility. The functional evolution of the WH2 domain has been approached by a combination of structural and biochemical methods, using thymosin beta4 (Tbeta4) and Ciboulot, a 3 beta-thymosin repeat protein from Drosophila as models. Ciboulot binds actin like Tbeta4 but promotes actin assembly like profilin. The first repeat of Ciboulot (D1) has the profilin function of the whole protein. The crystal structure of Ciboulot-actin shows that the major interaction with G-actin lies in the N-terminal amphipathic helix of D1. By point mutagenesis the sequestering activity of Tbeta4 can be changed into a profilin activity. ((1)H, (15)N)-NMR studies show that the functional switch from inhibition to promotion of actin assembly is linked to a change in the dynamics of interaction of the central and C-terminal regions of the WH2 domain with subdomains 1 and 2 of G-actin. Further systematic mutagenesis studies have been performed by engineering a series of chimeras of Ciboulot and Tbeta4. Proteins displaying either profilin function or enhanced sequestering activity compared to Tbeta4 have been characterized. The results provide insight into the structural basis for the regulation of the multiple functions of the WH2 domain.

Published
2007

20. Backbone chemical shift assignments of human 14-3-3σ

Author

Isabelle Landrieu, João Neves, Emmanuelle Boll, François-Xavier Cantrelle, Hamida Merzougui, Xavier Hanoulle, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), and Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
Gene isoform, Models, Molecular, 030303 biophysics, Chemical biology, Computational biology, Biochemistry, Protein Structure, Secondary, Protein–protein interaction, 03 medical and health sciences, Structural Biology, Interaction network, NMR resonance assignments, Humans, Amino Acid Sequence, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Phosphorylated proteins, Nuclear Magnetic Resonance, Biomolecular, 030304 developmental biology, 0303 health sciences, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Mechanism (biology), Drug discovery, Chemistry, Signal transducing adaptor protein, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Quantitative Biology - Biomolecules, 14-3-3 Proteins, Protein-Protein Interactions
Abstract

14-3-3 proteins are a group of seven dimeric adapter proteins that exert their biological function by interacting with hundreds of phosphorylated proteins, thus influencing their sub-cellular localization, activity or stability in the cell. Due to this remarkable interaction network, 14-3-3 proteins have been associated with several pathologies and the protein–protein interactions (PPIs) established with a number of partners are now considered promising drug targets. The activity of 14-3-3 proteins is often isoform specific and to our knowledge only one out of seven isoforms, 14-3-3 $$\zeta$$ , has been assigned. Despite the availability of the crystal structures of all seven isoforms of 14-3-3, the additional NMR assignments of 14-3-3 proteins are important for both biological mechanism studies and chemical biology approaches. Herein, we present a robust backbone assignment of 14-3-3σ, which will allow advances in the discovery of potential therapeutic compounds. This assignment is now being applied to the discovery of both inhibitors and stabilizers of 14-3-3 PPIs.

Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results