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40 results on '"MESH: Protease Inhibitors"'

1. Characterising proteolysis during SARS-CoV-2 infection identifies viral cleavage sites and cellular targets with therapeutic potential

Author

Jeanne Chiaravalli, Marius Walter, Edward Emmott, Emma Sierecki, Stacy Gellenoncourt, Philip Brownridge, Arturas Grauslys, Andrew R. Jones, Fabrice Agou, Charles S. Craik, Marco Vignuzzi, Yann Gambin, Patrick A. Eyers, Lisa A. Chakrabarti, Eric Verdin, Leonard A. Daly, Bjoern Meyer, Dominic P. Bryne, Claire E. Eyers, Populations virales et Pathogenèse - Viral Populations and Pathogenesis, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Criblage chémogénomique et biologique (Plateforme) - Chemogenomic and Biological Screening Platform (PF-CCB), Institut Pasteur [Paris] (IP), Virus et Immunité - Virus and immunity (CNRS-UMR3569), University of Liverpool, Buck Institute for Research on Aging, University of California [San Francisco] (UC San Francisco), University of California (UC), University of New South Wales [Sydney] (UNSW), This work was supported by the Laboratoire d’Excellence 'Integrative Biology of Emerging Infectious Diseases' (grant ANR-10-LABX-62-IBEID) to M.V. S.G. is the recipient of a MESR/Ecole Doctorale BioSPC ED562, Université de Paris fellowship. L.A.C. is supported by Institut Pasteur TASK FORCE SARS COV-2 (Tropicoro project), DIM ELICIT Region Ile-de-France, and ANRS. E.E. is supported by startup funding from the University of Liverpool, as well as a Wellcome Trust ISSF Interdisciplinary & Industry Award. E.E. is grateful for the support of GoFundMe donors for sponsoring SARS-CoV-2 research in his laboratory., and ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010)

Subjects
Proteomics, MESH: Virus Internalization, viruses, medicine.medical_treatment, General Physics and Astronomy, Virus Replication, MESH: Dipeptides, MESH: RNA, Small Interfering, 2.2 Factors relating to the physical environment, MESH: COVID-19, MESH: Animals, MESH: Myosin-Light-Chain Kinase, RNA, Small Interfering, Aetiology, skin and connective tissue diseases, Lung, MESH: Protease Inhibitors, MESH: Viral Proteases, Multidisciplinary, medicine.diagnostic_test, Viral Proteases, MESH: Proteomics, Proteases, Dipeptides, src-Family Kinases, Infectious Diseases, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Infection, Biotechnology, Proto-oncogene tyrosine-protein kinase Src, MESH: Antiviral Agents, MESH: Mutation, Science, Proteolysis, MESH: Proteolysis, Context (language use), Biology, Small Interfering, Antiviral Agents, Article, General Biochemistry, Genetics and Molecular Biology, Cell Line, Vaccine Related, Viral Proteins, Biodefense, medicine, Animals, Humans, Protease Inhibitors, MESH: SARS-CoV-2, Myosin-Light-Chain Kinase, MESH: Humans, Protease, SARS-CoV-2, Prevention, fungi, MESH: Virus Replication, COVID-19, MYLK, Pneumonia, General Chemistry, Virus Internalization, MESH: Viral Proteins, Virology, COVID-19 Drug Treatment, MESH: Cell Line, body regions, Emerging Infectious Diseases, Good Health and Well Being, MESH: src-Family Kinases, Viral replication, Mutation, RNA, Immunization
Abstract

SARS-CoV-2 is the causative agent behind the COVID-19 pandemic, responsible for over 170 million infections, and over 3.7 million deaths worldwide. Efforts to test, treat and vaccinate against this pathogen all benefit from an improved understanding of the basic biology of SARS-CoV-2. Both viral and cellular proteases play a crucial role in SARS-CoV-2 replication. Here, we study proteolytic cleavage of viral and cellular proteins in two cell line models of SARS-CoV-2 replication using mass spectrometry to identify protein neo-N-termini generated through protease activity. We identify previously unknown cleavage sites in multiple viral proteins, including major antigens S and N: the main targets for vaccine and antibody testing efforts. We discover significant increases in cellular cleavage events consistent with cleavage by SARS-CoV-2 main protease, and identify 14 potential high-confidence substrates of the main and papain-like proteases. We show that siRNA depletion of these cellular proteins inhibits SARS-CoV-2 replication, and that drugs targeting two of these proteins: the tyrosine kinase SRC and Ser/Thr kinase MYLK, show a dose-dependent reduction in SARS-CoV-2 titres. Overall, our study provides a powerful resource to understand proteolysis in the context of viral infection, and to inform the development of targeted strategies to inhibit SARS-CoV-2 and treat COVID-19., During SARS-CoV-2 replication, viral and cellular proteases play crucial roles and have been shown to be promising anti-viral targets. Here, Meyer et al. apply mass spectrometry to characterize the proteolytic cleavage profile of viral and cellular proteins in vitro.

Published
2021

2. Potency, safety, and pharmacokinetics of the NS3/4A protease inhibitor BI201335 in patients with chronic HCV genotype-1 infection

Author

Christian Trepo, Michael P. Manns, David Wright, Jose Luis Calleja, Peter W. White, Chan-Loi Yong, Stanislas Pol, George Kukolj, Jerry O. Stern, Marc Bourlière, Maurizio Bonacini, Thomas Berg, Wulf Otto Boecher, Yves Benhamou, Gerhard G. Steinmann, Joe Scherer, Department of Gastroenterology, Hepatology and Endocrinology (MHH), Hannover Medical School [Hannover] (MHH), Hôpital saint joseph, Service d' Hépato-gastroentérologie, Service d'Hépato-Gastro-Entérologie [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'hépatologie médicale [CHU Cochin], Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), California Pacific medical research center institute, Pacific medical center, Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), IFR Laennec (IL), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet [Saint-Étienne] (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of Psychiatry and Behavioural Neurosciences, McMaster University [Hamilton, Ontario]-Offord Centre for Child Studies, University Clinic, University Clinic, Leipzig, Hospital universitano Puerta de Hierro, hospital universitano puerta de hierro, Service d'Hépato-Gastroentérologie [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Pitié-Salpêtrière [APHP], CHU Cochin [AP-HP]-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Centre de Recherche en Cancérologie de Lyon (CRCL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of Gastroenterology, Hepatology and Endocrinology ( MHH ), Hannover Medical School [Hannover] ( MHH ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP], Institut Cochin ( UM3 (UMR 8104 / U1016) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP], Centre de Recherche en Cancérologie de Lyon ( CRCL ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), IFR Laennec ( IL ), Institut National de la Recherche Agronomique ( INRA ) -Université Claude Bernard Lyon 1 ( UCBL ), and Université de Lyon-Université de Lyon-Université Jean Monnet [Saint-Étienne] ( UJM ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
Male, Aminoisobutyric Acids, Hepacivirus, MESH : Viral Load, Viral Nonstructural Proteins, medicine.disease_cause, Gastroenterology, MESH : Hepacivirus, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, Polyethylene Glycols, MESH: Recombinant Proteins, chemistry.chemical_compound, 0302 clinical medicine, MESH : Thiazoles, Pegylated interferon, MESH : Female, MESH: Double-Blind Method, MESH: Hepacivirus, MESH : Viral Nonstructural Proteins, 0303 health sciences, MESH: Middle Aged, MESH: Protease Inhibitors, MESH: Drug Resistance, Viral, biology, MESH : Polyethylene Glycols, Intracellular Signaling Peptides and Proteins, MESH : Adult, Middle Aged, Viral Load, Hepatitis C, Recombinant Proteins, MESH : Antiviral Agents, 3. Good health, MESH : Drug Resistance, Viral, MESH: Oligopeptides, Quinolines, MESH : Interferon-alpha, Female, 030211 gastroenterology & hepatology, MESH : Carrier Proteins, MESH: Interferon-alpha, MESH: Viral Load, Oligopeptides, Viral load, medicine.drug, Pegylated Interferon Alfa, MESH : Oligopeptides, Adult, MESH: Antiviral Agents, medicine.medical_specialty, MESH : Protease Inhibitors, Proline, MESH : Recombinant Proteins, MESH : Male, Hepatitis C virus, MESH: Thiazoles, MESH: Carrier Proteins, [SDV.CAN]Life Sciences [q-bio]/Cancer, Interferon alpha-2, Antiviral Agents, 03 medical and health sciences, Double-Blind Method, Leucine, MESH: Ribavirin, Internal medicine, Drug Resistance, Viral, Ribavirin, medicine, MESH : Double-Blind Method, Humans, MESH : Middle Aged, Protease Inhibitors, Protease inhibitor (pharmacology), 030304 developmental biology, MESH: Hepatitis C, MESH: Humans, Hepatology, business.industry, MESH : Humans, Interferon-alpha, MESH: Adult, MESH : Hepatitis C, biology.organism_classification, MESH: Male, MESH : Ribavirin, Thiazoles, MESH: Polyethylene Glycols, chemistry, Faldaprevir, Immunology, MESH: Viral Nonstructural Proteins, Carrier Proteins, business, MESH: Female
Abstract

International audience; BACKGROUND & AIMS: BI201335 is a highly specific and potent HCV protease inhibitor. This multiple rising dose trial evaluated antiviral activity and safety in chronic HCV genotype-1 patients. METHODS: Thirty-four treatment-naïve patients were randomized to monotherapy with placebo or BI201335 at 20-240 mg once-daily for 14 days, followed by combination with pegylated interferon alfa/ribavirin (PegIFN/RBV) through Day 28. Nineteen treatment-experienced patients received 48-240 mg BI201335 once-daily with PegIFN/RBV for 28 days. HCV-RNA was measured with Roche COBAS TaqMan. RESULTS: In treatment-naïve patients, median maximal viral load (VL) reductions during 14-day monotherapy were -3.0, -3.6, -3.7, and -4.2 log(10) for the 20, 48, 120, and 240 mg groups. VL breakthroughs (≥1 log(10) from nadir) were seen in most patients on monotherapy and were caused by NS3/4A variants (R155K, D168V) conferring in vitro resistance to BI201335. Adding PegIFN/RBV at Days 15-28 led to continuous viral load reductions in most patients. In treatment-experienced patients, treatment with BI201335 and PegIFN/RBV achieved VL

Published
2011

3. A New Generation of SpecificTrypanosoma cruzi trans-Sialidase Inhibitors

Author

Stephen G. Withers, Alejandro Buschiazzo, Sabrina Buchini, University of British Columbia (UBC), Institut Pasteur de Montevideo, Réseau International des Instituts Pasteur (RIIP), and Canadian Institutes for Health ResearchSwiss National Science Foundation

Subjects
Models, Molecular, Stereochemistry, Trypanosoma cruzi, MESH: Molecular Structure, MESH: Glycoproteins, Neuraminidase, Crystallography, X-Ray, 010402 general chemistry, Sensitivity and Specificity, 01 natural sciences, Catalysis, Trans-sialidase, Hydrolase, [CHIM.CRIS]Chemical Sciences/Cristallography, Animals, Bioorganic chemistry, Protease Inhibitors, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, ComputingMilieux_MISCELLANEOUS, Glycoproteins, chemistry.chemical_classification, MESH: Protease Inhibitors, Molecular Structure, biology, MESH: Kinetics, 010405 organic chemistry, Chemistry, MESH: Neuraminidase, General Chemistry, General Medicine, biology.organism_classification, MESH: Crystallography, X-Ray, MESH: Sensitivity and Specificity, 0104 chemical sciences, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Kinetics, Enzyme, Biochemistry, MESH: Trypanosoma cruzi, MESH: Models, Molecular
Abstract

International audience

Published
2008

4. Naturally Occurring Resistance-Associated Variants of Hepatitis C Virus Protease Inhibitors in Poor Responders to Pegylated Interferon-Ribavirin

Author

Larrat, Sylvie, Vallet, Sophie, David-Tchouda, Sandra, Caporossi, Alban, Margier, Jennifer, Ramière, Christophe, Scholtès, Caroline, Haïm-Boukobza, Stéphanie, Roque-Afonso, Anne-Marie, Besse, Bernard, André-Garnier, Elisabeth, Mohamed, Sofiane, Halfon, Philippe, Pivert, Adeline, Leguillou-Guillemette, Hélène, Abravanel, Florence, Guivarch, Matthieu, Mackiewicz, Vincent, Lada, Olivier, Mourez, Thomas, Plantier, Jean-Christophe, Baazia, Yazid, Alain, Sophie, Hantz, Sébastien, Thibault, Vincent, Gaudy-Graffin, Catherine, Bouvet, Dorine, Mirand, Audrey, Henquell, Cécile, Gozlan, Joël, Lagathu, Gisèle, Pronier, Charlotte, Velay, Aurélie, SCHVOERER, Evelyne, Trimoulet, Pascale, Fleury, Hervé, Bouvier-Alias, Magali, BROCHOT, Etienne, Duverlie, Gilles, Maylin, Sarah, Gouriou, Stéphanie, Pawlotsky, Jean-Michel, Morand, Patrice, Plantier, Christophe, Tang, Y.-W., Unit for Virus Host-Cell Interactions [Grenoble] (UVHCI), Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Universitaire de Biodiversité et Ecologie Microbienne (LUBEM), Université de Brest (UBO), Centre d'Investigation Clinique - Innovation Technologique - INSERM - CHU de Grenoble (CIC-IT Grenoble (CIT803)), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Grenoble Alpes - UFR Médecine (UGA UFRM), Université Grenoble Alpes (UGA), Centre Hospitalier Universitaire [Grenoble] (CHU), Biologie cellulaire des infections virales – Cell biology of viral infection, Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Lyon (CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Laboratoire de virologie, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Paul Brousse, Hôtel-Dieu de Nantes, Alphabio Laboratory, Hôpital Européen [Fondation Ambroise Paré - Marseille], Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM), Centre de Physiopathologie Toulouse Purpan (ex IFR 30 et IFR 150) (CPTP), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UPS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpital Beaujon, Centre de recherche sur l'Inflammation (CRI), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), CHU Rouen, Normandie Université (NU), Service de bactériologie-Virologie-Hygiène [Avicenne], Université Paris 13 (UP13)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Avicenne, Université de Limoges (UNILIM), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, CHU Pontchaillou [Rennes], Morphogénèse et antigénicité du VIH et du virus des Hépatites (MAVIVH), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), Epidémiologie et pathogénie des infections à entérovirus (EPIE), Université d'Auvergne - Clermont-Ferrand I (UdA), Laboratoire Microorganismes : Génome et Environnement - Clermont Auvergne (LMGE), Université Clermont Auvergne (UCA)-Centre National de la Recherche Scientifique (CNRS), CHU Saint-Antoine [APHP], Virology Laboratory, General Hospital Papageorgiou, Service de Virologie [CHRU Nancy], Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy), Stress, Immunité, Pathogènes (SIMPA), Université de Lorraine (UL), Service de virologie et d'immunologie biologique, CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence Virus des hépatites B, C et Delta, Institut National de la Transfusion Sanguine [Paris] (INTS)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Unité de Virologie clinique et fondamentale EA 4294, CHU Amiens-Picardie, 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, Laboratoire de Virologie, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Laboratoire de virologie moléculaire et structurale, CHU Grenoble, Centre National de la Recherche Scientifique (CNRS)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Université Joseph Fourier - Grenoble 1 (UJF), Laboratoire de Parasitologie et Mycologiede [CHRU Brest], Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), Unité d'évaluation clinique et médico-économique [CHU Grenoble] (Département de la recherche clinique et de l'innovation), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Virologie [HCL-Hôpital de la Croix Rousse], Hôpital de la Croix-Rousse [CHU - HCL], Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL), Service de Virologie [Villejuif], Hôpital Paul Brousse, Université Paris-Sud - Paris 11 - Faculté de médecine (UP11 UFR Médecine), Université Paris-Sud - Paris 11 (UP11), Laboratoire de Virologie [CHU Nantes], Centre hospitalier universitaire de Nantes (CHU Nantes), ALPHABIO - Laboratoire de biologie médicale, Laboratoire de virologie [CHU Angers], PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM), Laboratoire de Virologie [CHU Purpan, Toulouse], Institut Fédératif de Biologie (IFB) - Hôpital Purpan, Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse], Hôpital Beaujon [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Laboratoire de virologie [Rouen], Hôpital Charles Nicolle [Rouen]-CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Département de microbiologie : Bactério, Virologie, Parasito, Hygiène, Université Paris 13 (UP13)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Avicenne [AP-HP], CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Service de bactériologie-virologie [Tours], Hôpital Bretonneau-Centre Hospitalier Régional Universitaire de Tours (CHRU Tours), Centre national de référence du VIH INSERM U966, Université de Tours (UT), CHU Clermont-Ferrand, Laboratoire de Virologie [CHU Saint-Antoine], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Laboratoire de Virologie [Rennes] = Virology [Rennes], Institut National de la Transfusion Sanguine [Paris] (INTS)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Laboratoire de bactériologie - hygiène [CHU Amiens], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)-Hopital Saint-Louis [AP-HP] (AP-HP), Unit for Virus Host-Cell Interactions [Grenoble] ( UVHCI ), Université Joseph Fourier - Grenoble 1 ( UJF ) -European Molecular Biology Laboratory [Grenoble] ( EMBL ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire Universitaire de Biodiversité et Ecologie Microbienne ( LUBEM ), Université de Brest ( UBO ), ThEMAS, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble ( TIMC-IMAG ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Institut d'Enseignement Supérieur et de Recherche en Alimentation, Santé Animale, Sciences Agronomiques et de l'Environnement ( VetAgro Sup ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Institut d'Enseignement Supérieur et de Recherche en Alimentation, Santé Animale, Sciences Agronomiques et de l'Environnement ( VetAgro Sup ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -CHU Grenoble, Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-CHU Grenoble, Pathogénèse et traitement de l'hépatite fulminante et du cancer du foie, Université Paris-Sud - Paris 11 ( UP11 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Grenoble Alpes - UFR Médecine ( UGA UFRM ), Université Grenoble Alpes ( UGA ), Internal Medicine and Infectious Diseases Department, European Hospital and Alphabio Laboratory, Centre Hospitalier Universitaire d'Angers ( CHU Angers ), PRES Université Nantes Angers Le Mans ( UNAM ), Laboratoire de Virologie [Toulouse], CHU Toulouse [Toulouse], Service d'hépatologie, Assistance publique - Hôpitaux de Paris (AP-HP) ( APHP ) -Université Paris Diderot - Paris 7 ( UPD7 ) -Hôpital Beaujon, Hôpital Bichat - Claude Bernard, Centre de recherche sur l'Inflammation ( CRI ), Université Paris Diderot - Paris 7 ( UPD7 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Recherche Biomédicale des Armées ( IRBA ), CHU Avicenne, Service de virologie, Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Assistance publique - Hôpitaux de Paris (AP-HP) ( APHP ) -CHU Pitié-Salpêtrière [APHP], Morphogénèse et antigénicité du VIH et du virus des Hépatites ( MAVIVH ), Université de Tours-Centre Hospitalier Régional Universitaire de Tours-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre de Calcul de l'IN2P3 ( CC-IN2P3 ), Institut National de Physique Nucléaire et de Physique des Particules du CNRS ( IN2P3 ) -Centre National de la Recherche Scientifique ( CNRS ), CHU Strasbourg, Virology Laboratory, Bordeaux University Hospital, Bordeaux, France, Microbiologie cellulaire et moléculaire et pathogénicité ( MCMP ), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique ( CNRS ), Faculté de Médecine, CHU Henri Mondor, Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Virologie [CHU Toulouse], Institut Fédératif de Biologie (IFB), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Pôle Biologie [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Hôpital Charles Nicolle [Rouen], Normandie Université (NU)-Normandie Université (NU)-CHU Rouen, Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Hôpital Bretonneau, SCHVOERER, Evelyne, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), CHU Saint-Antoine [AP-HP], Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Département de microbiologie : Bactério, Virologie, Parasito, Hygiène-Hôpital Charles Nicolle [Rouen]-CHU Rouen, and Université de Tours

Subjects
Male, Hepacivirus, Viral Nonstructural Proteins, medicine.disease_cause, Gastroenterology, Telaprevir, chemistry.chemical_compound, Pegylated interferon, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Hepacivirus, MESH: Treatment Outcome, MESH: Inhibitory Concentration 50, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Aged, education.field_of_study, MESH: Drug Resistance, Viral, MESH: Middle Aged, MESH: Protease Inhibitors, Hepatitis C, Middle Aged, 3. Good health, MESH: Hepatitis C, Chronic, Treatment Outcome, MESH: Oligopeptides, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Female, MESH: Interferon-alpha, Viral load, Oligopeptides, medicine.drug, Microbiology (medical), MESH: Antiviral Agents, Adult, medicine.medical_specialty, Proline, Hepatitis C virus, Population, Mutation, Missense, Antiviral Agents, Inhibitory Concentration 50, MESH: Ribavirin, Internal medicine, Boceprevir, Virology, Drug Resistance, Viral, Ribavirin, medicine, Humans, Protease Inhibitors, education, Aged, Retrospective Studies, MESH: Mutation, Missense, MESH: Humans, MESH: Proline, business.industry, Interferon-alpha, MESH: Adult, MESH: Retrospective Studies, Hepatitis C, Chronic, medicine.disease, MESH: Male, chemistry, Immunology, MESH: Viral Nonstructural Proteins, business, MESH: Female, [ SDV.BBM.BS ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

The pretherapeutic presence of protease inhibitor (PI) resistance-associated variants (RAVs) has not been shown to be predictive of triple-therapy outcomes in treatment-naive patients. However, they may influence the outcome in patients with less effective pegylated interferon (pegIFN)-ribavirin (RBV) backbones. Using hepatitis C virus (HCV) population sequence analysis, we retrospectively investigated the prevalence of baseline nonstructural 3 (NS3) RAVs in a multicenter cohort of poor IFN-RBV responders (i.e., prior null responders or patients with a viral load decrease of 3-fold shift in 50% inhibitory concentration (IC 50 ) for telaprevir or boceprevir, T54S was the most frequently detected mutation (3.9%), followed by A156T, R155K (0.7%), V36M, and V55A (0.35%). Mutations were more frequently found in patients infected with genotype 1a (7.5 to 23.6%) than 1b (3.3 to 19.8%) ( P = 0.03). No other sociodemographic or viroclinical characteristic was significantly associated with a higher prevalence of RAVs. No obvious effect of baseline RAVs on viral load was observed. In this cohort of poor responders to IFN-RBV, no link was found with a sustained virological response to triple therapy, regardless of the algorithm used for the detection of mutations. Based on a cross-study comparison, baseline RAVs are not more frequent in poor IFN-RBV responders than in treatment-naive patients and, even in these difficult-to-treat patients, this study demonstrates no impact on treatment outcome, arguing against resistance analysis prior to treatment.

Published
2015

5. Effect of doxycycline on sulfur mustard-induced respiratory lesions in guinea pigs.: Doxycycline Prevents SM-Induced Respiratory Lesions

Author

Marie-Pia d'Ortho, Stéphane Galiacy, Laurent Taysse, Séraphin Delamanche, Emmanuelle Planus, Jean-Henri Calvet, Christophe Guignabert, Physiopathologie et Thérapeutiques Respiratoires, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10, Centre d'Etudes du Bouchet, Délégation Générale pour l'Armement, Service de physiologie, explorations fonctionnelles [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), and This work was funded by the DGA and the Institut National de la Sante et de la Recherche Medicale (France).

Subjects
Male, MESH: Inflammation, Gelatinases, Physiology, MESH: Chemical Warfare Agents, Respiratory System, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Matrix metalloproteinase, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, chemistry.chemical_compound, 0302 clinical medicine, vesicant agents, Gelatinase, MESH: Animals, MESH: Gelatinases, Chemical Warfare Agents, MESH: Doxycycline, MESH: Respiratory System, Doxycycline, 0303 health sciences, MESH: Protease Inhibitors, Sulfur mustard, Anti-Bacterial Agents, 3. Good health, medicine.anatomical_structure, MESH: Epithelial Cells, [SDV.TOX]Life Sciences [q-bio]/Toxicology, 030220 oncology & carcinogenesis, medicine.drug, Pulmonary and Respiratory Medicine, Guinea Pigs, protease inhibitors, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, MESH: Guinea Pigs, Microbiology, Guinea pig, 03 medical and health sciences, [SDV.MHEP.CSC]Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, MESH: Anti-Bacterial Agents, Physiology (medical), Mustard Gas, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], medicine, Animals, Zymography, 030304 developmental biology, Inflammation, Tissue Inhibitor of Metalloproteinase-2, Tissue Inhibitor of Metalloproteinase-1, doxycycline, blistering agents, Epithelial Cells, Cell Biology, Molecular biology, MESH: Male, MESH: Mustard Gas, MESH: Tissue Inhibitor of Metalloproteinase-2, MESH: Tissue Inhibitor of Metalloproteinase-1, chemistry, proteases, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Respiratory tract
Abstract

9 pages; International audience; Respiratory tract lesions induced by the chemical warfare agent sulfur mustard (SM) are characterized by epithelial damages associated with inflammatory cell infiltration. Here we evaluated the imbalance between gelatinase and tissue inhibitors of metalloproteinases (TIMPs), and we tested pretreatment with the protease inhibitor doxycycline. Guinea pigs were intoxicated intratracheally with SM and evaluated 24 h after exposure. Matrix metalloproteinase (MMP) gelatinase activity of bronchial lavage (BL) fluid from SM-exposed guinea pigs was high compared with controls, as shown by both zymography and biotinylated substrate degradation, whereas TIMP-1 and -2 levels by immunoblotting were similar. Extensive areas of lysis were evidenced by in situ zymography, indicating imbalance between gelatinases and inhibitors towards net proteolytic activity. Doxycycline pretreatment resulted in 1) decreased gelatinase activity (zymography, free gelatinase activity assay, and in situ zymography); 2) decreased inflammation (BL fluid cellularity and protein level); and 3) dramatic decrease in histological epithelial lesions. Our results suggest inadequate levels of TIMP to counteract increased gelatinase activity and further support a role for MMP gelatinases in SM-induced respiratory lesions. They also suggest that doxycycline may hold promise as a therapeutic tool.

Published
2005

6. Aminopeptidase-N/CD13 (EC 3.4.11.2) inhibitors: Chemistry, biological evaluations, and therapeutic prospects

Author

Brigitte Bauvois, Daniel Dauzonne, Signalisation cellulaire, dynamique circulatoire et athérosclérose précoce (SCDCAP), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Hydroxamic Acids, natural inhibitor, Angiogenesis, medicine.medical_treatment, Pharmacology, Hydroxamic Acids, 01 natural sciences, MESH: Tyrosine, 0302 clinical medicine, MESH: Curcumin, Drug Discovery, MESH: Animals, Disulfides, Receptor, Neprilysin, 0303 health sciences, aminopeptidase, MESH: Protease Inhibitors, MESH: Neprilysin, Chemistry, MESH: Antigens, CD13, General Medicine, Transmembrane protein, 3. Good health, 030220 oncology & carcinogenesis, MESH: Oligopeptides, Molecular Medicine, medicine.symptom, Pentacyclic Triterpenes, Oligopeptides, MESH: Neovascularization, Physiologic, hormones, hormone substitutes, and hormone antagonists, Cell type, bestatin, Curcumin, animal structures, Neovascularization, Physiologic, Inflammation, ectoenzyme, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, CD13 Antigens, Article, 03 medical and health sciences, Immune system, Leucine, MESH: Cell Proliferation, synthetic inhibitor, medicine, cancer, Animals, Humans, Protease Inhibitors, MESH: Disulfides, Secretion, Betulinic Acid, Cell Proliferation, 030304 developmental biology, MESH: Humans, Protease, 010405 organic chemistry, Cell growth, MESH: Triterpenes, Triterpenes, 0104 chemical sciences, MESH: Leucine, inflammation, Tyrosine
Abstract

Aminopeptidase N (APN)/CD13 (EC 3.4.11.2) is a transmembrane protease present in a wide variety of human tissues and cell types (endothelial, epithelial, fibroblast, leukocyte). APN/CD13 expression is dysregulated in inflammatory diseases and in cancers (solid and hematologic tumors). APN/CD13 serves as a receptor for coronaviruses. Natural and synthetic inhibitors of APN activity have been characterized. These inhibitors have revealed that APN is able to modulate bioactive peptide responses (pain management, vasopressin release) and to influence immune functions and major biological events (cell proliferation, secretion, invasion, angiogenesis). Therefore, inhibition of APN/CD13 may lead to the development of anti‐cancer and anti‐inflammatory drugs. This review provides an update on the biological and pharmacological profiles of known natural and synthetic APN inhibitors. Current status on their potential use as therapeutic agents is discussed with regard to toxicity and specificity. © 2005 Wiley Periodicals, Inc. Med Res Rev

Published
2005

7. Antiinfective therapy with a small molecule inhibitor of Staphylococcus aureus sortase

Author

Ruihan Zhang, Olaf Schneewind, Shaynoor Dramsi, Feifei Chen, Lu Zhou, Hualiang Jiang, Ya-Ting Wang, Jiafei Li, Cai-Guang Yang, Jie Zhang, Lefu Lan, Cheng Luo, Shouzhe Gong, Hongchuan Liu, Kongkai Zhu, Shanghai Institute of Materia Medica - Chinese Academy of Sciences [Shanghai], University of Chicago, Biologie des Bactéries pathogènes à Gram-positif, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Fudan University [Shanghai], This work was supported by National Science and Technology Major Project 'Key New Drug Creation and Manufacturing Program' Grant 2013ZX09507-004, National Natural Science Foundation of China Grants 91313303, 20972173, and 81230076, and Hi-Tech Research and Development Program of China Grants 2012AA020302 and 2012AA020301. Work on sortase in the laboratory of O.S. is supported by National Institute of Allergy and Infectious Diseases Grant AI038897. Computation resources were partially supported by the Computer Network Information Center, Chinese Academy of Sciences, and Shanghai Supercomputing Center., and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Models, Molecular, [SDV]Life Sciences [q-bio], Antibiotics, medicine.disease_cause, computational screening, Anti-Infective Agents, Sortase, MESH: Staphylococcus aureus, antivirulence, MESH: Animals, MESH: Bacterial Proteins, 0303 health sciences, Mice, Inbred BALB C, MESH: Microbial Sensitivity Tests, Multidisciplinary, MESH: Protease Inhibitors, MESH: Peptides, Clostridium difficile, Biological Sciences, Aminoacyltransferases, 3. Good health, Cysteine Endopeptidases, Staphylococcus aureus, Sortase A, nosocomial infection, Female, MESH: Streptococcus pyogenes, MESH: Biocatalysis, MESH: Models, Molecular, medicine.drug_class, Streptococcus pyogenes, MESH: Anti-Infective Agents, MESH: Mice, Inbred BALB C, Microbial Sensitivity Tests, MESH: Thiadiazoles, Biology, LPXTG motif, Microbiology, Sepsis, Small Molecule Libraries, 03 medical and health sciences, Bacterial Proteins, MESH: Small Molecule Libraries, MESH: Aminoacyltransferases, Thiadiazoles, medicine, Animals, Protease Inhibitors, 030304 developmental biology, 030306 microbiology, medicine.disease, Virology, Bacteremia, Biocatalysis, Peptides, MESH: Female, MESH: Cysteine Endopeptidases
Abstract

International audience; Methicillin-resistant Staphylococcus aureus (MRSA) is the most frequent cause of hospital-acquired infection, which manifests as surgical site infections, bacteremia, and sepsis. Due to drug-resistance, prophylaxis of MRSA infection with antibiotics frequently fails or incites nosocomial diseases such as Clostridium difficile infection. Sortase A is a transpeptidase that anchors surface proteins in the envelope of S. aureus, and sortase mutants are unable to cause bacteremia or sepsis in mice. Here we used virtual screening and optimization of inhibitor structure to identify 3-(4-pyridinyl)-6-(2-sodiumsulfonatephenyl)[1,2,4]triazolo[3,4-b][1,3,4]thiadiazole and related compounds, which block sortase activity in vitro and in vivo. Sortase inhibitors do not affect in vitro staphylococcal growth yet protect mice against lethal S. aureus bacteremia. Thus, sortase inhibitors may be useful as antiinfective therapy to prevent hospital-acquired S. aureus infection in high-risk patients without the side effects of antibiotics.

Published
2014

8. Morbillivirus Infection of the Mouse Central Nervous System Induces Region-Specific Upregulation of MMPs and TIMPs Correlated to Inflammatory Cytokine Expression

Author

Hideo Akaoka, Seng-Thuon Khuth, Olivier Verlaeten, Axel Pagenstecher, Arlette Bernard, Pascale Giraudon, Marie-Françoise Belin, Centre de recherche en neurosciences de Lyon (CRNL), Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Université Jean Monnet [Saint-Étienne] (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV]Life Sciences [q-bio], Matrix metalloproteinase, Virus Replication, MESH: Dogs, Extracellular matrix, Mice, MESH: Gene Expression Regulation, Viral, 0302 clinical medicine, MESH: Up-Regulation, MESH: Animals, Distemper Virus, Canine, Regulation of gene expression, MESH: Cytokines, 0303 health sciences, MESH: Protease Inhibitors, biology, Brain, Metalloendopeptidases, Up-Regulation, Cell biology, medicine.anatomical_structure, Cytokines, Female, Tumor necrosis factor alpha, Gene Expression Regulation, Viral, MESH: Distemper, Immunology, Central nervous system, MESH: Metalloendopeptidases, Microbiology, MESH: Brain, 03 medical and health sciences, Dogs, Immune system, Downregulation and upregulation, Virology, MESH: Distemper Virus, Canine, medicine, Animals, Humans, Protease Inhibitors, RNA, Messenger, Distemper, Interleukin 6, MESH: Mice, MESH: RNA, Messenger, 030304 developmental biology, Tissue Inhibitor of Metalloproteinase-3, Tissue Inhibitor of Metalloproteinase-1, MESH: Humans, MESH: Virus Replication, Disease Models, Animal, MESH: Tissue Inhibitor of Metalloproteinase-1, MESH: Tissue Inhibitor of Metalloproteinase-3, Insect Science, biology.protein, Pathogenesis and Immunity, MESH: Disease Models, Animal, MESH: Female, 030217 neurology & neurosurgery
Abstract

Viral infection of the central nervous system (CNS) can result in perturbation of cell-to-cell communication involving the extracellular matrix (ECM). ECM integrity is maintained by a dynamic balance between the synthesis and proteolysis of its components, mainly as a result of the action of matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs). An MMP/TIMP imbalance may be critical in triggering neurological disorders, in particular in virally induced neural disorders. In the present study, a mouse model of brain infection using a neurotropic strain of canine distemper virus (CDV) was used to study the effect of CNS infection on the MMP/TIMP balance and cytokine expression. CDV replicates almost exclusively in neurons and has a unique pattern of expression (cortex, hypothalamus, monoaminergic nuclei, hippocampus, and spinal cord). Here we show that although several mouse brain structures were infected, they exhibited a differential pattern in terms of MMP, TIMP, and cytokine expression, exemplified by (i) a large increase in pro-MMP9 levels, in particular in the hippocampus, which occurred mainly in neurons and was associated with in situ gelatinolytic activity, (ii) specific and significant upregulation of MT1-MMP mRNA expression in the cortex and hypothalamus, (iii) an MMP/TIMP imbalance, suggested by the upregulation of TIMP-1 mRNA in the cortex, hippocampus, and hypothalamus and of TIMP-3 mRNA in the cortex, and (iv) a concomitant region-specific large increase in expression of Th1-like cytokines, such as gamma interferon, tumor necrosis factor alpha, and interleukin 6 (IL-6), contrasting with weaker induction of Th2-like cytokines, such as IL-4 and IL-10. These data indicate that an MMP/TIMP imbalance in specific brain structures, which is tightly associated with a local inflammatory process as shown by the presence of immune infiltrating cells, differentially impairs CNS integrity and may contribute to the multiplicity of late neurological disorders observed in this viral mouse model.

Published
2001

9. Evidence by Site-Directed Mutagenesis That Arginine 203 of Thermolysin and Arginine 717 of Neprilysin (Neutral Endopeptidase) Play Equivalent Critical Roles in Substrate Hydrolysis and Inhibitor Binding

Author

Emmanuel Ruffet, Marie-Claude Fournie-Zaluski, Bernard P. Roques, Michael J. O’Donohue, Serge Antonczak, Cynthia Marie-Claire, Ann Beaumont, Pharmacochimie moléculaire et structurale, 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 ) -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 ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Médicaments : Dynamique Intracellulaire et Architecture Nucléaire ( MéDIAN ), Université de Reims Champagne-Ardenne ( URCA ) -Centre National de la Recherche Scientifique ( CNRS ), 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)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Médicaments : Dynamique Intracellulaire et Architecture Nucléaire (MéDIAN), Université de Reims Champagne-Ardenne (URCA)-Centre National de la Recherche Scientifique (CNRS), Fractionnement des AgroRessources et Environnement - UMR-A 614 (FARE), Université de Reims Champagne-Ardenne (URCA)-SFR Condorcet, Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS)-Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), 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)-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)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Marie-Claire, Cynthia

Subjects
Models, Molecular, MESH : Molecular Sequence Data, Arginine, MESH : DNA, Complementary, MESH: Amino Acid Sequence, Biochemistry, Substrate Specificity, MESH: Recombinant Proteins, MESH : Arginine, Site-directed mutagenesis, MESH : Thiorphan, Neprilysin, MESH : Neprilysin, chemistry.chemical_classification, 0303 health sciences, MESH: Protease Inhibitors, MESH : Substrate Specificity, biology, MESH: Neprilysin, Chemistry, MESH : Amino Acid Sequence, Hydrolysis, 030302 biochemistry & molecular biology, MESH: Arginine, Glycopeptides, MESH : Bacillus subtilis, Recombinant Proteins, MESH : Mutagenesis, Site-Directed, MESH: Mutagenesis, Site-Directed, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Hydrolysis, MESH: Models, Molecular, Bacillus subtilis, MESH : Glycopeptides, Thiorphan, MESH : Protease Inhibitors, DNA, Complementary, MESH : Recombinant Proteins, MESH : Models, Molecular, Molecular Sequence Data, Thermolysin, MESH: Binding, Competitive, Binding, Competitive, MESH: Thermolysin, 03 medical and health sciences, MESH : Hydrolysis, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Protease Inhibitors, Amino Acid Sequence, Enzyme kinetics, MESH : Thermolysin, MESH: Glycopeptides, 030304 developmental biology, MESH: Molecular Sequence Data, MESH : Binding, Competitive, Mutagenesis, Active site, MESH: Thiorphan, MESH: DNA, Complementary, MESH: Bacillus subtilis, Enzyme, Mutagenesis, Site-Directed, biology.protein, MESH: Substrate Specificity
Abstract

52 ref. 2 tables 4 graph.; International audience; Neprilysin (neutral endopeptidase-24.11, EC 3.4.24.11) is a mammalian zinc-endopeptidase involved in the degradation of biologically active peptides. Although no atomic structure is available for this enzyme, site-directed mutagenesis studies have shown that its active site resembles closely that of the bacterial zinc-endopeptidase, thermolysin (EC 3.4.24.27). One active site residue of thermolysin, Arg-203, is involved in inhibitor binding by forming hydrogen bonds with the carbonyl group of a residue in the P1 position and also participates in a hydrogen bond network involving Asp-170. Sequence alignment data shows that Arg-717 of neprilysin could play a similar role to Arg-203 of thermolysin. This was investigated by site-directed mutagenesis with Arg-203 of thermolysin and Arg-717 of neprilysin being replaced by methionine residues. This led, in both cases, to decreases in kcat/Km values, of 122-fold for neprilysin and 2300-fold for thermolysin, essentially due to changes in kcat. The Ki values of several inhibitors were also increased for the mutated enzymes. In addition, the replacement of Asp-170 of thermolysin by Ala residue resulted in a decrease in kcat/Km of 220-fold. The results, coupled with a molecular modeling study, suggest that Arg-717 of neprilysin corresponds to Arg-203 of thermolysin and that in both enzymes a hydrogen bond network exists, involving His-142, Asp-170, and Arg-203 in thermolysin and His-583, Asp-650, and Arg-717 in neprilysin, which is crucial for hydrolytic activity.

Published
1997

10. Nevirapine-raltegravir combination, an NRTI and PI/r sparing regimen, as maintenance antiretroviral therapy in virologically suppressed HIV-1-infected patients

Author

Elisabeth André-Garnier, Virginie Ferré, Dominique Bettinger, Catherine Chirouze, Bruno Hoen, Gilles Peytavin, François Raffi, Patrice Muret, Clotilde Allavena, Véronique Reliquet, Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Service des maladies infectieuses et tropicales, Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Saint-Jacques, Service de maladies infectieuses et tropicales [Nantes], Université de Nantes ( UN ) -Hôtel-Dieu-Centre hospitalier universitaire de Nantes ( CHU Nantes ), Infection, Antimicrobiens, Modélisation, Evolution ( IAME ), Université Paris 13 ( UP13 ) -Université Paris Diderot - Paris 7 ( UPD7 ) -Université Sorbonne Paris Cité ( USPC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre hospitalier universitaire de Nantes ( CHU Nantes ), CHU Pointe-à-Pitre/Abymes [Guadeloupe], Maladies infectieuses et tropicales, Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Hôpital Saint-Jacques, Université de Nantes (UN)-Hôtel-Dieu-Centre hospitalier universitaire de Nantes (CHU Nantes), Infection, Anti-microbiens, Modélisation, Evolution (IAME (UMR_S_1137 / U1137)), Université Paris 13 (UP13)-Université Paris Diderot - Paris 7 (UPD7)-Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre hospitalier universitaire de Nantes (CHU Nantes), services de maladies infectieuses, CHU J Minjoz, Besançon, Ministère de la santé, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Ingénierie et biologie cellulaire et tissulaire (IBCT (ex IFR133)), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Service de pharmacie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-AP-HP - Hôpital Bichat - Claude Bernard [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC ( HOTE GREFFON ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Ingénierie et biologie cellulaire et tissulaire ( IBCT (ex IFR133) ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), and Assistance publique - Hôpitaux de Paris (AP-HP)-AP-HP - Hôpital Bichat - Claude Bernard [Paris]

Subjects
Male, MESH : Pyrrolidinones, MESH: CD4 Lymphocyte Count, MESH : Retrospective Studies, MESH : Viral Load, HIV Infections, MESH: Antiretroviral Therapy, Highly Active, Raltegravir Potassium, MESH: HIV-1, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Risk Factors, [ SDV.MP ] Life Sciences [q-bio]/Microbiology and Parasitology, Risk Factors, Antiretroviral Therapy, Highly Active, Pharmacology (medical), MESH : Female, 030212 general & internal medicine, MESH: Nevirapine, MESH : Reverse Transcriptase Inhibitors, MESH: Treatment Outcome, MESH : Drug Substitution, 0303 health sciences, MESH: Middle Aged, MESH: Protease Inhibitors, Reverse-transcriptase inhibitor, Drug Substitution, virus diseases, MESH: Follow-Up Studies, MESH: HIV Infections, Middle Aged, Viral Load, MESH : Risk Factors, Pyrrolidinones, 3. Good health, [ SDV.MHEP.MI ] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Drug Combinations, Infectious Diseases, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, MESH : Drug Combinations, Treatment Outcome, Reverse Transcriptase Inhibitors, Female, MESH: Viral Load, Viral load, MESH : HIV-1, medicine.drug, Nevirapine, MESH : Protease Inhibitors, MESH : Male, MESH : Nevirapine, MESH : Treatment Outcome, MESH: Pyrrolidinones, 03 medical and health sciences, medicine, MESH : CD4 Lymphocyte Count, MESH : HIV Infections, Humans, Protease inhibitor (pharmacology), MESH : Middle Aged, Protease Inhibitors, Retrospective Studies, Pharmacology, MESH: Drug Combinations, MESH: Humans, 030306 microbiology, business.industry, MESH : Humans, MESH: Retrospective Studies, MESH : Follow-Up Studies, Raltegravir, MESH : Antiretroviral Therapy, Highly Active, Virology, MESH: Drug Substitution, MESH: Male, CD4 Lymphocyte Count, Regimen, HIV-1, Ritonavir, MESH: Reverse Transcriptase Inhibitors, business, MESH: Female, Follow-Up Studies
Abstract

Background Nucleoside reverse transcriptase inhibitor (NRTI) and protease inhibitor (PI)/ritonavir sparing regimens may be useful to some HIV-infected patients. Nevirapine (NVP) and raltegravir (RAL) are both potent antiretrovirals with good long-term safety profiles. Methods We retrospectively identified from our electronic database all patients with HIV RNA6 months on an NVP-containing regimen and no prior exposure to integrase strand transfer inhibitors who were switched to RAL plus NVP. Data was collected for 36 months or until discontinuation of RAL plus NVP for any reason. Results A total of 39 patients (30 male) were included in this analysis. Median duration of prior antiretroviral therapy was 14 years (IQR 10–17) and median duration with plasma HIV-1 RNAConclusions In patients with prolonged HIV-1 RNA

Published
2013

11. Multicenter quality control of hepatitis C virus protease inhibitor resistance genotyping

Author

Evelyne Schvoerer, Elisabeth André-Garnier, Joël Gozlan, Etienne Brochot, Sylvie Larrat, Sébastien Hantz, Catherine Gaudy-Graffin, Hélène Le Guillou-Guillemette, Nelly Magnat, Vincent Mackiewicz, Syria Laperche, Jean-Christophe Plantier, Philippe Colson, Françoise Lunel-Fabiani, Patrick Soussan, Gisèle Lagathu, Christopher Payan, Jacques Izopet, Sylvain Rosec, S. Gouriou, Anne-Marie Roque-Afonso, Patrice Morand, Caroline Scholtes, Adeline Pivert, Sophie Vallet, Florence Legrand-Abravanel, Vincent Thibault, Sarah Maylin, Audrey Mirand, Stéphane Chevaliez, Jean-Michel Pawlotsky, Charlotte Charpentier, Valérie Giordanengo, Cécile Henquell, Françoise Bouchardeau, Lina Aguilera, Pascale Trimoulet, Laboratoire Universitaire de Biodiversité et Ecologie Microbienne (LUBEM), Université de Brest (UBO), Laboratoire de virologie moléculaire et structurale (LVMS), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence Virus des hépatites B, C et Delta, Institut National de la Transfusion Sanguine [Paris] (INTS)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Laboratoire de virologie, Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM), Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Virologie [Rennes] = Virology [Rennes], CHU Pontchaillou [Rennes], Service de virologie [CHU Pitié-Salpêtrière], Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Laboratoire de Virologie, CHU Strasbourg, Virus, pseudo-virus: Morphogénèse et Antigénicité, Université de Tours-EA3856, CHU Bordeaux [Bordeaux], Laboratoire de Virologie [CHU Amiens], CHU Amiens-Picardie, Unité de Virologie clinique et fondamentale (UVCF), CHU Amiens-Picardie-Université de Picardie Jules Verne (UPJV), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, Services des Maladies Infectieuses et Tropicales [CHU Saint-Antoine], CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Paul Brousse, Carcinogenèse Hépatique et Virologie Moléculaire, Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de virologie [Rouen], Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Département de microbiologie : Bactério, Virologie, Parasito, Hygiène-Hôpital Charles Nicolle [Rouen]-CHU Rouen, Normandie Université (NU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-AP-HP - Hôpital Bichat - Claude Bernard [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR48, INSB-INSB-Centre National de la Recherche Scientifique (CNRS), Centre d'Investigation Clinique (CIC - Brest), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Virologie [Purpan], CHU Toulouse [Toulouse]-Institut Fédératif de Biologie (IFB) - Hôpital Purpan, Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse], Université de Tours (UT)-EA3856, Hôpital Charles Nicolle [Rouen]-CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Département de microbiologie : Bactério, Virologie, Parasito, Hygiène, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-IFR10, Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS), Service de virologie, Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-CHU Pontchaillou [Rennes], Sorbonne Université-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université, Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie, Service des maladies infectieuses et tropicales [CHU Saint-Antoine], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université-Sorbonne Université, Université Nice Sophia Antipolis (1965 - 2019) (UNS), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Hôpital Charles Nicolle [Rouen], CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-CHU Rouen, Institut des sciences biologiques (INSB-CNRS)-Institut des sciences biologiques (INSB-CNRS)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Virologie [CHU Toulouse], Institut Fédératif de Biologie (IFB), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Pôle Biologie [CHU Toulouse], and Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)

Subjects
Microbiology (medical), Quality Control, MESH: Antiviral Agents, MESH: Sequence Analysis, DNA, MESH: Mutation, Genotype, Hepatitis C virus, Hepacivirus, [SDV]Life Sciences [q-bio], MESH: Quality Control, MESH: Amino Acid Sequence, Viral Nonstructural Proteins, MESH: Base Sequence, medicine.disease_cause, Antiviral Agents, MESH: Genotype, 03 medical and health sciences, 0302 clinical medicine, Virology, Drug Resistance, Viral, medicine, Protease inhibitor (pharmacology), Protease Inhibitors, MESH: Hepacivirus, Amino Acid Sequence, Genotyping, 0303 health sciences, NS3, MESH: Drug Resistance, Viral, MESH: Protease Inhibitors, biology, Base Sequence, 030306 microbiology, Inhibitor resistance, virus diseases, Sequence Analysis, DNA, biology.organism_classification, 3. Good health, Hcv protease, Mutation, MESH: Viral Nonstructural Proteins, 030211 gastroenterology & hepatology
Abstract

Hepatitis C virus (HCV) protease inhibitor resistance-associated substitutions are selected during triple-therapy breakthrough. This multicenter quality control study evaluated the expertise of 23 French laboratories in HCV protease inhibitor resistance genotyping. A panel of 12 well-defined blinded samples comprising two wild-type HCV strains, nine transcripts from synthetic NS3 mutant samples or from clinical strains, and one HCV RNA-negative sample was provided to the participating laboratories. The results showed that any laboratory with expertise in sequencing techniques should be able to provide reliable HCV protease inhibitor resistance genotyping. Only a 0.7% error rate was reported for the amino acid sites studied. The accuracy of substitution identification ranged from 75% to 100%, depending on the laboratory. Incorrect results were mainly related to the methodology used. The results could be improved by changing the primers and modifying the process in order to avoid cross-contamination. This study underlines the value of quality control programs for viral resistance genotyping, which is required prior to launching observational collaborative multicenter studies on HCV resistance to direct-acting antiviral agents.

Published
2013

12. Group X secreted phospholipase A2 proenzyme is matured by a furin-like proprotein convertase and releases arachidonic acid inside of human HEK293 cells

Author

Christine Payré, Khaoula Chargui, Rob C. Oslund, Sabine Scarzello, Michael H. Gelb, Hiromi, Gérard Lambeau, Anne Sophie Dabert-Gay, Dominique Douguet, Ikram Jemel, Institut de pharmacologie moléculaire et cellulaire (IPMC), Centre National de la Recherche Scientifique (CNRS)-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), Department of Chemistry, University of Washington [Seattle], and Department of Biochemistry [Washington ]

Subjects
MESH: Group X Phospholipases A2, MESH: Mutation, medicine.medical_treatment, Amino Acid Motifs, Biochemistry, 03 medical and health sciences, Mice, MESH: Amino Acid Motifs, MESH: Enzyme Precursors, 0302 clinical medicine, Phospholipase A2, medicine, Animals, Group X Phospholipases A2, Humans, Secretion, Protease Inhibitors, MESH: Animals, Protein precursor, Molecular Biology, Furin, MESH: Mice, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Enzyme Precursors, Protease, Arachidonic Acid, MESH: Protease Inhibitors, MESH: Humans, biology, Cell Biology, Proprotein convertase, Lipids, Amino acid, MESH: Arachidonic Acid, HEK293 Cells, chemistry, 030220 oncology & carcinogenesis, MESH: HEK293 Cells, Mutation, biology.protein, MESH: Proprotein Convertases, [SDV.IMM]Life Sciences [q-bio]/Immunology, Proprotein Convertases
Abstract

International audience; Among mammalian secreted phospholipases A(2) (sPLA(2)s), group X sPLA(2) has the most potent hydrolyzing activity toward phosphatidylcholine and is involved in arachidonic acid (AA) release. Group X sPLA(2) is produced as a proenzyme and contains a short propeptide of 11 amino acids ending with a dibasic motif, suggesting cleavage by proprotein convertases. Although the removal of this propeptide is clearly required for enzymatic activity, the cellular location and the protease(s) involved in proenzyme conversion are unknown. Here we have analyzed the maturation of group X sPLA(2) in HEK293 cells, which have been extensively used to analyze sPLA(2)-induced AA release. Using recombinant mouse (PromGX) and human (ProhGX) proenzymes; HEK293 cells transfected with cDNAs coding for full-length ProhGX, PromGX, and propeptide mutants; and various permeable and non-permeable sPLA(2) inhibitors and protease inhibitors, we demonstrate that group X sPLA(2) is mainly converted intracellularly and releases AA before externalization from the cell. Most strikingly, the exogenous proenzyme does not elicit AA release, whereas the transfected proenzyme does elicit AA release in a way insensitive to non-permeable sPLA(2) inhibitors. In transfected cells, a permeable proprotein convertase inhibitor, but not a non-permeable one, prevents group X sPLA(2) maturation and partially blocks AA release. Mutations at the dibasic motif of the propeptide indicate that the last basic residue is required and sufficient for efficient maturation and AA release. All together, these results argue for the intracellular maturation of group X proenzyme in HEK293 cells by a furin-like proprotein convertase, leading to intracellular release of AA during secretion.

Published
2011

13. Hepatitis C virus (HCV) protease variability and anti-HCV protease inhibitor resistance in HIV/HCV-coinfected patients

Author

Trimoulet, P., Belzunce, C., Faure, M., Wittkop, L., Reigadas, S., Dupon, M., Ragnaud, J.-M., Fleury, H., Neau, Didier, Service de virologie et d'immunologie biologique, CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, Laboratoire Réactions et Génie des Procédés (LRGP), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), and CHU Pellegrin

Subjects
MESH: Antiviral Agents, Adult, Male, MESH: Mutation, Genotype, MESH: Peptide Hydrolases, HIV Infections, Hepacivirus, Viral Nonstructural Proteins, Antiviral Agents, MESH: Genotype, Drug Resistance, Viral, Humans, MESH: Hepacivirus, Protease Inhibitors, MESH: Hepatitis C, MESH: Middle Aged, MESH: Protease Inhibitors, MESH: Drug Resistance, Viral, MESH: Humans, Coinfection, virus diseases, MESH: Adult, MESH: HIV Infections, Middle Aged, Hepatitis C, digestive system diseases, MESH: Male, MESH: Coinfection, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Mutation, MESH: Viral Nonstructural Proteins, Female, MESH: Female, Peptide Hydrolases
Abstract

International audience; OBJECTIVES: Data on the natural selection of isolates harbouring mutations within the NS3 protease, conferring resistance to hepatitis C virus (HCV) protease inhibitors (PIs), are limited for HIV/HCV-coinfected patients. The aim of this study was to describe the natural prevalence of mutations conferring resistance to HCV PIs in HIV/HCV-coinfected patients compared with HCV-monoinfected patients. METHODS: The natural prevalences of HCV PI resistance mutations in 120 sequences from HIV/HCV-coinfected patients (58 genotype 1a, 18 genotype 1b and 44 genotype 4) and 501 sequences from HCV-monoinfected patients (476 genotype 1 and 25 genotype 4), retrieved from GenBank as a control group, were compared. RESULTS: Of 76 sequences from HIV/HCV genotype 1-coinfected patients, six (7.9%) showed amino acid substitutions associated with HCV PI resistance (V36L, n=1; V36M, n=2; T54S, n=2; R155K, n=1). In 31 of 476 (6.5%) HCV genotype 1 sequences retrieved from the GenBank database, HCV PI resistance mutations were found. The difference was not statistically significant (P=0.6). All of the sequences from HIV/HCV genotype 4-coinfected patients and those retrieved from the GenBank database had amino acid changes at position 36 (V36L). CONCLUSION: Our study suggests that the natural prevalence of strains resistant to HCV PIs does not differ between HCV-monoinfected and HIV/HCV-coinfected patients. Further studies on larger cohorts are needed to confirm these findings and to evaluate the impact of these mutations in clinical practice.

Published
2011

14. Danoprevir, an HCV NS3/4A protease inhibitor, improves insulin sensitivity in patients with genotype 1 chronic hepatitis C

Author

Michel Vidaud, Rami Moucari, Stefan Zeuzem, Yves Benhamou, Dominique Guyader, Bill Bradford, Patrice Couzigou, Nicole Forestier, Hélène Voitot, Patrick Marcellin, Scott D. Seiwert, Dominique Larrey, Centre de recherche biomédicale Bichat-Beaujon (CRB3), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Johann Wolfgang Goethe University Medical Center, Microenvironnement et remodelage, Centre Hospitalier Universitaire [Rennes]-Foie, métabolismes et cancer, Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Département d'hépatologie et de gastroentérologie, Université Bordeaux Segalen - Bordeaux 2-CHU Bordeaux [Bordeaux]-Hôpital Haut-Lévêque [CHU Bordeaux], CHU Bordeaux [Bordeaux], Service d'Hépato-Gastro-Entérologie [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service de Biochimie et Génétique Moléculaire, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)-Hôpital Beaujon [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), InterMune Laboratory, Brisbane, California, USA., Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Service d'Hépato-Gastroentérologie [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Pitié-Salpêtrière [APHP], and Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Université Paris Diderot - Paris 7 (UPD7)-Hôpital Beaujon

Subjects
Cyclopropanes, Male, medicine.medical_treatment, MESH: Sulfonamides, Hepacivirus, Isoindoles, Viral Nonstructural Proteins, medicine.disease_cause, Gastroenterology, MESH: Genotype, chemistry.chemical_compound, 0302 clinical medicine, MESH: Hepacivirus, Sulfonamides, 0303 health sciences, MESH: Middle Aged, MESH: Protease Inhibitors, Danoprevir, virus diseases, Hepatitis C, Middle Aged, Viral Load, 3. Good health, MESH: Hepatitis C, Chronic, MESH: Insulin Resistance, MESH: RNA, Viral, RNA, Viral, Female, 030211 gastroenterology & hepatology, MESH: Viral Load, Viral load, Adult, MESH: Antiviral Agents, medicine.medical_specialty, Genotype, Lactams, Proline, Lactams, Macrocyclic, Hepatitis C virus, Antiviral Agents, 03 medical and health sciences, Insulin resistance, Internal medicine, medicine, Humans, Protease Inhibitors, Protease inhibitor (pharmacology), 030304 developmental biology, MESH: Humans, MESH: Lactams, business.industry, Ribavirin, Insulin, MESH: Adult, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Hepatitis C, Chronic, medicine.disease, MESH: Male, chemistry, Immunology, MESH: Viral Nonstructural Proteins, Insulin Resistance, business, MESH: Female
Abstract

International audience; BACKGROUND/AIM: Insulin resistance (IR) is a major predictor of treatment failure in patients with hepatitis C virus (HCV) infection treated with peginterferon/ribavirin. The aim of this study was to evaluate the short-term effect of an HCV protease inhibitor monotherapy on IR in parallel with an antiviral effect. PATIENTS/METHODS: In a phase 1b placebo-controlled study, four cohorts of treatment-naïve patients with genotype 1 HCV received danoprevir (ITMN-191/RG7227), a protease inhibitor, or placebo (8/2 patients in each cohort respectively) in a gelatin capsule every 12 h (100, 200 mg) or 8 h (100, 200 mg) for 14 days. A fifth cohort including prior non-responders to peginterferon/ribavirin was similarly randomised to receive placebo or 300 mg danoprevir every 12 h. IR was assessed with the homeostasis model (HOMA-IR) at baseline and days 7, 14 and 15. RESULTS: Serum HCV-RNA and HOMA-IR correlated significantly (Spearman rho=0.379, p

Published
2010

15. Blastocystis legumain is localized on the cell surface, and specific inhibition of its activity implicates a pro-survival role for the enzyme

Author

Michaël Roussel, Catherine Texier, Binhui Wu, Kevin S. W. Tan, Jing Yin, Department of Microbiology, Yong Loo Lin School of Medicine, Laboratoire Microorganismes : Génome et Environnement (LMGE), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Centre National de la Recherche Scientifique (CNRS)-Université d'Auvergne - Clermont-Ferrand I (UdA), Infectious Disease Program, Life Sciences Institute, National University of Singapore, Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Centre National de la Recherche Scientifique (CNRS), and National University of Singapore (NUS)

Subjects
MESH: Cell Death, MESH: Hydrogen-Ion Concentration, MESH: Annexin A5, Cell, MESH: Amino Acid Sequence, Biochemistry, Substrate Specificity, law.invention, MESH: Antibodies, Monoclonal, Mice, 0302 clinical medicine, law, MESH: Animals, Annexin A5, MESH: In Situ Nick-End Labeling, 0303 health sciences, MESH: Protease Inhibitors, Cell Death, MESH: Escherichia coli, Antibodies, Monoclonal, MESH: Phosphatidylserines, Hydrogen-Ion Concentration, Cysteine Endopeptidases, Protein Transport, MESH: Cattle, medicine.anatomical_structure, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, MESH: Cell Survival, 030220 oncology & carcinogenesis, Recombinant DNA, DNA fragmentation, MESH: Blastocystis, Programmed cell death, MESH: Protein Transport, MESH: Rats, Cell Survival, Molecular Sequence Data, DNA Fragmentation, Phosphatidylserines, Biology, Legumain, 03 medical and health sciences, Enzyme activator, Molecular Basis of Cell and Developmental Biology, Escherichia coli, In Situ Nick-End Labeling, medicine, Animals, Humans, MESH: DNA Fragmentation, Protease Inhibitors, Amino Acid Sequence, Molecular Biology, MESH: Mice, 030304 developmental biology, Blastocystis, MESH: Humans, MESH: Molecular Sequence Data, Cell Biology, biology.organism_classification, Molecular biology, Rats, biology.protein, Cattle, MESH: Substrate Specificity, MESH: Cysteine Endopeptidases
Abstract

International audience; Programmed cell death (PCD) is crucial for cellular growth and development in multicellular organisms. Although distinct PCD features have been described for unicellular eukaryotes, homology searches have failed to reveal clear PCD-related orthologues among these organisms. Our previous studies revealed that a surface-reactive monoclonal antibody (mAb) 1D5 could induce multiple PCD pathways in the protozoan Blastocystis. In this study, we identified, by two-dimensional gel electrophoresis and mass spectrometry, the target of mAb 1D5 as a surface-localized legumain, an asparagine endopeptidase that is usually found in lysosomal/acidic compartments of other organisms. Recombinant Blastocystis legumain displayed biphasic pH optima in substrate assays, with peaks at pH 4 and 7.5. Activity of Blastocystis legumain was greatly inhibited by the legumain-specific inhibitor carbobenzyloxy-Ala-Ala-AAsn-epoxycarboxylate ethyl ester (APE-RR) (where AAsn is aza-asparagine) and moderately inhibited by mAb 1D5, cystatin, and caspase-1 inhibitor. Interestingly, inhibition of legumain activity induced PCD in Blastocystis, observed by increased externalization of phosphatidylserine residues and in situ DNA fragmentation. In contrast to plants, in which legumains have been shown to play a pro-death role, legumain appears to display a pro-survival role in Blastocystis.

Published
2010

16. NAD(P)H quinone-oxydoreductase 1 protects eukaryotic translation initiation factor 4GI from degradation by the proteasome

Author

Rodica Anesia, Stéphane Pyronnet, Philippe Pierre, Corinne Bousquet, Bertrand Fabre, Christiane Susini, Amandine Alard, Catherine Marboeuf, Simon, Marie Francoise, Institut de médecine moléculaire de Rangueil (I2MR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre d'Immunologie de Marseille - Luminy (CIML), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Chirurgie Générale et Digestive [Rangueil], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IFR150-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), CHU Toulouse [Toulouse]-Hôpital de Rangueil, CHU Toulouse [Toulouse], and Université de Toulouse (UT)-Université de Toulouse (UT)- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Small interfering RNA, MESH: NAD(P)H Dehydrogenase (Quinone), Mice, Peptide Initiation Factors, Chlorocebus aethiops, NAD(P)H Dehydrogenase (Quinone), Protein biosynthesis, MESH: Animals, MESH: Peptide Initiation Factors, MESH: Peptide Fragments, 0303 health sciences, MESH: Protease Inhibitors, MESH: Oxidative Stress, Protein Stability, MESH: Proteasome Endopeptidase Complex, 030302 biochemistry & molecular biology, Articles, Transfection, MESH: Eukaryotic Initiation Factor-4G, Biochemistry, MESH: Protein Biosynthesis, Proteasome Inhibitors, Protein Binding, Proteasome Endopeptidase Complex, MESH: NADPH Dehydrogenase, Biology, Cell Line, 03 medical and health sciences, Eukaryotic translation, MESH: Protein Stability, Animals, Humans, Initiation factor, MESH: Protein Binding, Protease Inhibitors, Molecular Biology, MESH: Mice, 030304 developmental biology, MESH: Humans, NADPH Dehydrogenase, Cell Biology, MESH: Cercopithecus aethiops, Peptide Fragments, MESH: Cell Line, Oxidative Stress, Proteasome, Protein Biosynthesis, NAD+ kinase, Eukaryotic Initiation Factor-4G
Abstract

International audience; The eukaryotic translation initiation factor 4GI (eIF4GI) serves as a central adapter in cap-binding complex assembly. Although eIF4GI has been shown to be sensitive to proteasomal degradation, how the eIF4GI steady-state level is controlled remains unknown. Here, we show that eIF4GI exists in a complex with NAD(P)H quinone-oxydoreductase 1 (NQO1) in cell extracts. Treatment of cells with dicumarol (dicoumarol), a pharmacological inhibitor of NQO1 known to preclude NQO1 binding to its protein partners, provokes eIF4GI degradation by the proteasome. Consistently, the eIF4GI steady-state level also diminishes upon the silencing of NQO1 (by transfection with small interfering RNA), while eIF4GI accumulates upon the overexpression of NQO1 (by transfection with cDNA). We further reveal that treatment of cells with dicumarol frees eIF4GI from mRNA translation initiation complexes due to strong activation of its natural competitor, the translational repressor 4E-BP1. As a consequence of cap-binding complex dissociation and eIF4GI degradation, protein synthesis is dramatically inhibited. Finally, we show that the regulation of eIF4GI stability by the proteasome may be prominent under oxidative stress. Our findings assign NQO1 an original role in the regulation of mRNA translation via the control of eIF4GI stability by the proteasome.

Published
2010

17. HIV-1 integrase-hydrolyzing antibodies from sera of HIV-infected patients

Author

Georgy A. Nevinsky, Vincent Parissi, Svetlana V. Baranova, Marie-Line Andreola, Marina A. Kharitonova, Valentina N. Buneva, Christina Calmels, Ludmila P. Sizyakina, Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), and Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, Leupeptins, Antibodies, Catalytic, HIV Infections, HIV Integrase, Biochemistry, Chromatography, Affinity, MESH: Pepstatins, Sepharose, Pepstatins, Sulfones, chemistry.chemical_classification, 0303 health sciences, MESH: Protease Inhibitors, biology, Hydrolysis, 030302 biochemistry & molecular biology, MESH: Sulfones, MESH: Chromatography, Gel, General Medicine, MESH: HIV Infections, MESH: Chromatography, Affinity, 3. Good health, Integrase, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, MESH: Young Adult, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, Female, Antibody, MESH: Hydrolysis, Adult, Proteases, MESH: Enzyme Activation, Adolescent, 03 medical and health sciences, Young Adult, Immune system, Antigen, Humans, Protease Inhibitors, 030304 developmental biology, MESH: Adolescent, MESH: Humans, Autoantibody, MESH: Adult, MESH: Leupeptins, Virology, MESH: Male, Enzyme Activation, Enzyme, chemistry, biology.protein, MESH: HIV Integrase, MESH: Female, MESH: Antibodies, Catalytic, MESH: Electrophoresis, Polyacrylamide Gel
Abstract

International audience; Autoantibodies with enzymic activities (abzymes) are a distinctive feature of autoimmune diseases. It was interesting whether Abs from patients with viral diseases can hydrolyze viral proteins. Electrophoretically and immunologically homogeneous IgGs were isolated from sera of AIDS patients by chromatography on several affinity sorbents. We present evidence showing that 89.5% IgGs purified from the sera of HIV-infected patients using several affinity resins including Sepharose with immobilized integrase specifically hydrolyze only HIV integrase (IN) but not many other tested proteins. Several rigid criteria have been applied to show that the IN-hydrolyzing activity is an intrinsic property of AIDS IgGs but not from healthy donors. Similar to autoimmune proteolytic abzymes, IN-hydrolyzing IgGs from some patients were inhibited by specific inhibitors of serine and metal-dependent proteases but a significant inhibition of the activity by specific inhibitors of acidic- and thiol-like proteases was observed for the first time. Although HIV infection leads to formation of Abs to many viral and human antigens, no possible biological role for most of them is known. Since anti-IN IgG can efficiently hydrolyze IN, a positive role of abzymes in counteracting the infection cannot be excluded. In addition, detection of IN-hydrolyzing activity can be useful for diagnostic purposes and for estimation of the immune status in AIDS patients.

Published
2009

18. Plasma citrulline is a biomarker of enterocyte mass and an indicator of parenteral nutrition in HIV-infected patients

Author

Luc Cynober, Nathalie Neveux, Pascal Crenn, Jean Claude Melchior, Pierre de Truchis, Tatiana Galperine, Crenn, Pascal, Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Département de médecine, Hôpital Raymond Poincaré [AP-HP]-Université Paris Descartes - Paris 5 (UPD5), Processus d’adaptation métaboliques et nouvelles stratégies nutritionnelles (EA 2498), Université Paris Descartes - Paris 5 (UPD5), Service de biochimie et de génétique moléculaire [CHU Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université de Versailles Saint-Quentin-en-Yvelines ( UVSQ ), AP-HP Hôpital Raymond Poincaré [Garches]-Université Paris Descartes - Paris 5 ( UPD5 ), Processus d’adaptation métaboliques et nouvelles stratégies nutritionnelles ( EA 2498 ), Université Paris Descartes - Paris 5 ( UPD5 ), and Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP]

Subjects
Parenteral Nutrition, 030309 nutrition & dietetics, [ SDV.AEN ] Life Sciences [q-bio]/Food and Nutrition, MESH : Prospective Studies, Medicine (miscellaneous), MESH : Diarrhea, HIV Infections, Gastroenterology, Body Mass Index, MESH: HIV-1, chemistry.chemical_compound, MESH: Enterocytes, 0302 clinical medicine, Citrulline, Enteropathy, Prospective Studies, 0303 health sciences, MESH : AIDS-Related Opportunistic Infections, Nutrition and Dietetics, MESH: Middle Aged, MESH: Protease Inhibitors, MESH: AIDS-Related Opportunistic Infections, MESH: HIV Infections, Middle Aged, MESH : Adult, 3. Good health, MESH : Enterocytes, Diarrhea, MESH: Diarrhea, C-Reactive Protein, 030220 oncology & carcinogenesis, Biomarker (medicine), MESH : Parenteral Nutrition, medicine.symptom, Lipodystrophy, Viral load, MESH : HIV-1, Adult, medicine.medical_specialty, MESH : Protease Inhibitors, MESH: Serum Albumin, Context (language use), Biology, MESH : Serum Albumin, MESH: Body Mass Index, 03 medical and health sciences, Internal medicine, MESH: C-Reactive Protein, medicine, MESH : HIV Infections, Humans, Protease Inhibitors, MESH : Middle Aged, Serum Albumin, MESH: Humans, AIDS-Related Opportunistic Infections, MESH : Humans, MESH: Biological Markers, MESH : Citrulline, MESH: Adult, MESH : C-Reactive Protein, medicine.disease, MESH: Prospective Studies, MESH : Biological Markers, MESH : Body Mass Index, [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, Enterocytes, Parenteral nutrition, chemistry, Immunology, HIV-1, MESH: Parenteral Nutrition, MESH: Citrulline, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Biomarkers
Abstract

International audience; BACKGROUND: Plasma citrulline is a biomarker of enterocyte mass and function in humans. OBJECTIVE: We evaluated citrulline in the reemerging context of diarrhea in HIV-infected patients receiving highly active antiretroviral therapy. DESIGN: This study prospectively measured citrulline in 6 groups of HIV-1 patients (n = 115): 1) undetectable viral load without chronic diarrhea (a; n = 40) and with protease inhibitor-associated toxic chronic diarrhea (b; n = 26), 2) detectable viral load and CD4 > 200/mm(3) without (a; n = 6) and with (b; n = 11) chronic diarrhea, and 3) detectable viral load and CD4 10 micromol/L; P < 0.05). Citrulline correlated positively with albumin (P < 0.01) and BMI (P < 0.05) and negatively with C-reactive protein (P < 0.01). When antiinfectious and nutritional therapies were successful (n = 18 of 25), citrulline normalized in 2-12 wk. Neither chronic hepatic or pancreatic disease nor lipodystrophy and the metabolic syndrome affected citrulline. Compared with control subjects (38 +/- 8 micromol/L), patients without chronic diarrhea (groups 1a, 2a, and 3a) had normal citrulline concentrations (36 +/- 6 micromol/L). CONCLUSIONS: Plasma citrulline is a reliable biomarker of enterocyte functional mass in HIV patients. Citrulline does not allow the etiologic diagnosis of enteropathy, but it can discriminate between protease inhibitor toxic diarrhea and infectious enteropathy and quantify the functional consequences, which makes it an objective tool for indicating the need for parenteral nutrition.

Published
2009

19. A new role for complement C3: regulation of antigen processing through an inhibitory activity

Author

Nicole Lefebvre, Patrice N. Marche, Christian L. Villiers, Marie-Bernadette Villiers, François Cretin, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Biochimie et biophysique des systèmes intégrés (BBSI), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Grenoble Alpes (UGA), and Duperray, Alain

Subjects
MESH: Down-Regulation, Substrate Specificity, Mice, 0302 clinical medicine, MESH: Animals, 0303 health sciences, Immunity, Cellular, Mice, Inbred BALB C, MESH: Protease Inhibitors, U937 cell, medicine.diagnostic_test, Antigen processing, MESH: Antigen-Presenting Cells, MESH: Complement C3, Complement C3, U937 Cells, Acquired immune system, 3. Good health, Biochemistry, [SDV.IMM]Life Sciences [q-bio]/Immunology, Proteolysis, Immunology, Antigen presentation, Complement, MESH: Mice, Inbred BALB C, Antigen-Presenting Cells, Down-Regulation, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, MESH: U937 Cells, MESH: Immunity, Cellular, 03 medical and health sciences, Immune system, Antigen, Tetanus Toxin, medicine, Animals, Humans, Protease Inhibitors, MESH: Tetanus Toxin, Antigen-presenting cell, C3, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, MESH: Mice, 030304 developmental biology, MESH: Humans, Macrophages, MESH: Antigen Presentation, MESH: Substrate Specificity, Lysosomes, 030215 immunology, MESH: Lysosomes
Abstract

International audience; Increasing evidence underlines the involvement of complement component C3 in the establishment of acquired immunity which appears to play a complex role and to act at different levels. As antigen proteolysis by antigen presenting cells is a key event in the control of antigen presentation efficiency, and consequently in the quality of the immune response, we investigated whether C3 could modulate this step. Our results demonstrate for the first time that C3 can interfere with antigen proteolysis: (i) proteolysis of tetanus toxin (TT) by the lysosomal fraction from a human monocytic cell line (U937) is impaired in the presence of C3, (ii) this effect is C3-specific and involves the C3c fragment of the protein, (iii) C3c is effective even after disulfide disruption, but none of its three constitutive peptides is individually accountable for this inhibitory effect and (iv) the target-protease(s) exhibit(s) a serine-protease activity. The physiological relevance of our results is demonstrated by experiments showing a subcellular colocalisation of TT and C3 after their uptake by U937 and the reduction of TT proteolysis once internalised together with C3. These results highlight a novel role for C3 that broadens its capacity to modulate acquired immune response.

Published
2008

20. NFkappaB-dependent control of BACE1 promoter transactivation by Abeta42

Author

Jean Sevalle, Virginie Buggia-Prevot, Frédéric Checler, Steffen Rossner, Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS), Department of Neurochemistry, Universität Leipzig [Leipzig], and Fondation pour la Recherche Médicale

Subjects
MESH: NF-kappa B, Endogeny, IκB kinase, Biochemistry, Transactivation, Amyloid beta-Protein Precursor, Mice, MESH: Protein Structure, Tertiary, 0302 clinical medicine, MESH: Amyloid beta-Protein Precursor, Aspartic Acid Endopeptidases, MESH: Animals, 0303 health sciences, MESH: Protease Inhibitors, MESH: Aspartic Endopeptidases, NF-kappa B, Presenilins, MESH: Presenilins, I-kappa B Kinase, MESH: Response Elements, Transcriptional Activation, Proteases, MESH: Enzyme Activation, MESH: Rats, MESH: Mice, Transgenic, Transgene, Mice, Transgenic, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Response Elements, Presenilin, Cell Line, 03 medical and health sciences, Alzheimer Disease, Complementary DNA, mental disorders, Animals, Humans, Protease Inhibitors, MESH: I-kappa B Kinase, Molecular Biology, MESH: Mice, 030304 developmental biology, MESH: Humans, HEK 293 cells, Cell Biology, Fibroblasts, Molecular biology, Protein Structure, Tertiary, Rats, MESH: Cell Line, Enzyme Activation, MESH: Amyloid Precursor Protein Secretases, MESH: Trans-Activation (Genetics), MESH: Fibroblasts, Amyloid Precursor Protein Secretases, 030217 neurology & neurosurgery, MESH: Alzheimer Disease
Abstract

Beta-amyloid (Abeta) peptides that accumulate in Alzheimer disease are generated from the beta-amyloid precursor protein (betaAPP) by cleavages by beta-secretase BACE1 and by presenilin-dependent gamma-secretase activities. Very few data document a putative cross-talk between these proteases and the regulatory mechanisms underlying such interaction. We show that presenilin deficiency lowers BACE1 maturation and affects both BACE1 activity and promoter transactivation. The specific gamma-secretase inhibitor DFK167 triggers the decrease of BACE1 activity in wild-type but not in presenilin-deficient fibroblasts. This decrease is also elicited by catalytically inactive gamma-secretase. The overexpression of APP intracellular domain (AICD), the gamma/epsilon-secretase-derived C-terminal product of beta-amyloid precursor protein, does not modulate BACE1 activity or promoter transactivation in fibroblasts and does not alter BACE1 expression in AICD transgenic brains of mice. A DFK167-sensitive increase of BACE1 activity is observed in cells overexpressing APPepsilon (the N-terminal product of betaAPP generated by epsilon-secretase cleavage harboring the Abeta domain but lacking the AICD sequence), suggesting that the production of Abeta could account for the modulation of BACE1. Accordingly, we show that HEK293 cells overexpressing wild-type betaAPP exhibit a DFK167-sensitive increase in BACE1 promoter transactivation that is increased by the Abeta-potentiating Swedish mutation. This effect was mimicked by exogenous application of Abeta42 but not Abeta40 or by transient transfection of cDNA encoding Abeta42 sequence. The IkappaB kinase inhibitor BMS345541 prevents Abeta-induced BACE1 promoter transactivation suggesting that NFkappaB could mediate this Abeta-associated phenotype. Accordingly, the overexpression of wild-type or Swedish mutated betaAPP does not modify the transactivation of BACE1 promoter constructs lacking NFkappaB-responsive element. Furthermore, APP/beta-amyloid precursor protein-like protein deficiency does not affect BACE1 activity and expression. Overall, these data suggest that physiological levels of endogenous Abeta are not sufficient per se to modulate BACE1 promoter transactivation but that exacerbated Abeta production linked to wild-type or Swedish mutated betaAPP overexpression modulates BACE1 promoter transactivation and activity via an NFkappaB-dependent pathway.

Published
2008

21. Epigallocatechin-3-gallate (EGCG) inhibits the migratory behavior of tumor bronchial epithelial cells

Author

Christine Terryn, Béatrice Nawrocki-Raby, S. Hazgui, Arnaud Bonnomet, Myriam Polette, Magali Milliot, Jérôme Cutrona, Jean-Marie Zahm, Philippe Birembaut, Plasticité de l'épithélium respiratoire dans les conditions normales et pathologiques - UMR-S 903 (PERPMP), SFR CAP Santé (Champagne-Ardenne Picardie Santé), Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Centre Hospitalier Universitaire de Reims (CHU Reims)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Reims Champagne-Ardenne (URCA), Biomolécules : interactions moléculaires, cellulaires et cellules-matrice extracellulaire (BIMCCME), Université de Reims Champagne-Ardenne (URCA)-Centre Hospitalier Universitaire de Reims (CHU Reims)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'histologie, cytologie, biologie cellulaire et moléculaire Pol Bouin, Université de Reims Champagne-Ardenne (URCA)-Centre Hospitalier Universitaire de Reims (CHU Reims), InCa, SPLF, ARC, Zahm, Jean-Marie, Université de Reims Champagne-Ardenne (URCA)-Centre Hospitalier Universitaire de Reims (CHU Reims)-Institut National de la Santé et de la Recherche Médicale (INSERM)-SFR CAP Santé (Champagne-Ardenne Picardie Santé), and Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)

Subjects
MESH: Cell Death, Time Factors, Matrix metalloproteinase inhibitor, Cell, Cell Culture Techniques, Vimentin, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, Catechin, MESH: Dose-Response Relationship, Drug, 3D cell culture, 0302 clinical medicine, Cell Movement, MESH: Reverse Transcriptase Polymerase Chain Reaction, heterocyclic compounds, MESH: Cell Movement, MESH: Bronchial Neoplasms, 0303 health sciences, MESH: Protease Inhibitors, Microscopy, Video, biology, Cell Death, Reverse Transcriptase Polymerase Chain Reaction, MESH: Gene Expression Regulation, Enzymologic, Bronchial Neoplasms, food and beverages, Cell migration, MESH: Gene Expression Regulation, Neoplastic, 3. Good health, Cell biology, Gene Expression Regulation, Neoplastic, medicine.anatomical_structure, MESH: Epithelial Cells, 030220 oncology & carcinogenesis, Matrix Metalloproteinase 2, Electrophoresis, Polyacrylamide Gel, Pulmonary and Respiratory Medicine, Programmed cell death, MESH: Cell Line, Tumor, MESH: Catechin, MESH: Antineoplastic Agents, Phytogenic, MESH: Imaging, Three-Dimensional, Matrix Metalloproteinase Inhibitors, complex mixtures, Gene Expression Regulation, Enzymologic, 03 medical and health sciences, Imaging, Three-Dimensional, Cell Line, Tumor, medicine, Humans, Neoplasm Invasiveness, Protease Inhibitors, RNA, Messenger, MESH: RNA, Messenger, 030304 developmental biology, lcsh:RC705-779, MESH: Cell Culture Techniques, MESH: Humans, Dose-Response Relationship, Drug, Cell growth, Research, MESH: Time Factors, Epithelial Cells, MESH: Neoplasm Invasiveness, lcsh:Diseases of the respiratory system, Antineoplastic Agents, Phytogenic, MESH: Matrix Metalloproteinase 2, MESH: Microscopy, Video, Cell culture, biology.protein, [SDV.MHEP.PSR] Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, MESH: Vimentin, sense organs, MESH: Electrophoresis, Polyacrylamide Gel
Abstract

Background Many studies associated the main polyphenolic constituent of green tea, (-)-Epigallocatechin-3-gallate (EGCG), with inhibition of cancers, invasion and metastasis. To date, most of the studies have focused on the effect of EGCG on cell proliferation or death. Since cell migration is an important mechanism involved in tumor invasion, the aim of the present work was to target another approach of the therapeutic effect of EGCG, by investigating its effect on the cell migratory behavior. Methods The effect of EGCG (at concentrations lower than 10 μg/ml) on the migration speed of invasive cells was assessed by using 2D and 3D models of cell culture. We also studied the effects of EGCG on proteinases expression by RT-PCR analysis. By immunocytochemistry, we analyzed alterations of vimentin organization in presence of different concentrations of EGCG. Results We observed that EGCG had an inhibitory effect of cell migration in 2D and 3D cell culture models. EGCG also inhibited MMP-2 mRNA and protein expression and altered the intermediate filaments of vimentin. Conclusion Taken together, our results demonstrate that EGCG is able to inhibit the migration of bronchial tumor cells and could therefore be an attractive candidate to treat tumor invasion and cell migration.

Published
2008

22. Cathepsin D Overexpressed by Cancer Cells Can Enhance Apoptosis-dependent Chemo-sensitivity Independently of Its Catalytic Activity

Author

Emmanuelle Liaudet-Coopman, Mélanie Beaujouin, Le Ster, Yves, controle de la progression des cancers hormonaux-dependants, Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM), and This work was supported by Institut National de la Santé et de la Recherche Médicale, University of Montpellier I, Association pour la Recherche sur le Cancer (grant 3344, E Liaudet-Coopman) and Ligue Nationale contre le Cancer who provided a fellowship for Mélanie Beaujouin.

Subjects
Programmed cell death, MESH: Antineoplastic Agents, Phytogenic, MESH: Cathepsin D, Cathepsin D, Apoptosis, [SDV.CAN]Life Sciences [q-bio]/Cancer, Caspase 3, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH: Caspase 9, Catalysis, Article, MESH: Pepstatins, chemistry.chemical_compound, Cytosol, [SDV.CAN] Life Sciences [q-bio]/Cancer, MESH: Cytosol, Neoplasms, Pepstatins, MESH: Caspase 3, Tumor Cells, Cultured, Humans, Protease Inhibitors, MESH: Neoplasms, MESH: Tumor Cells, Cultured, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Etoposide, MESH: Etoposide, Caspase-9, MESH: Humans, MESH: Protease Inhibitors, biology, MESH: Apoptosis, Cytochrome c, Cytochromes c, MESH: Cytochromes c, MESH: Catalysis, Antineoplastic Agents, Phytogenic, Molecular biology, Caspase 9, MESH: Drug Resistance, Neoplasm, Cell biology, chemistry, Drug Resistance, Neoplasm, Cancer cell, biology.protein, Pepstatin
Abstract

The aspartic protease cathepsin D (cath-D) is a key mediator of induced-apoptosis and its proteolytic activity has been generally involved in this event. During apoptosis, cath-D is translocated to the cytosol. Since cath-D is one of the lysosomal enzymes which requires a more acidic pH to be proteolytically-active relative to the cysteine lysosomal enzymes such as cath-B and -L, it is therefore open to question whether cytosolic cath-D might be able to cleave substrate(s) implicated in the apoptotic cascade. Here we have investigated the role of wild-type cath-D and its proteolytically-inactive counterpart over-expressed by 3Y1-Ad12 cancer cells during chemotherapeutic-induced cytotoxicity and apoptosis, as well as the relevance of cath-D catalytic function. We demonstrate that wild-type or mutated catalytically-inactive cath-D strongly enhances chemo-sensitivity and apoptotic response to etoposide. Both wild-type and mutated inactive cath-D are translocated to the cytosol, increasing the release of cytochrome c, the activation of caspases-9 and -3 and the induction of a caspase-dependent apoptosis. In addition, pre-treatment of cells with the aspartic protease inhibitor, pepstatin A, does not prevent apoptosis. Interestingly therefore, the stimulatory effect of cath-D on cell death is independent of its catalytic activity. Overall, our results imply that cytosolic cath-D stimulates apoptotic pathways by interacting with a member of the apoptotic machinery rather than by cleaving specific substrate(s).

Published
2008

23. Inhibition of renin angiotensin system decreases renal protein oxidative damage in diabetic rats

Author

Stefan Schäfer, Jordi Boada, Jean-Pierre Girolami, Reinald Pamplona, Mariona Jové, Wolfgang Linz, Marie Buléon, Manuel Portero-Otin, Hugo Gonzalo, Ivan Tack, Simon, Marie Francoise, Department of Experimental Medicine, University of Lleida-IRBLLEIDA-PCiTA, Institut de médecine moléculaire de Rangueil (I2MR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM), Therapeutic Department Cardiovascular Research, and Sanofi Aventis

Subjects
Glycation End Products, Advanced, Angiotensin receptor, MESH: Rats, Zucker, Angiotensin-Converting Enzyme Inhibitors, Kidney, Protein oxidation, Biochemistry, Renin-Angiotensin System, chemistry.chemical_compound, Ramipril, MESH: Renin-Angiotensin System, MESH: Losartan, Diabetic Nephropathies, MESH: Proteins, MESH: Animals, MESH: Ramipril, MESH: Oxidative Stress, MESH: Protease Inhibitors, biology, Chemistry, MESH: Angiotensin-Converting Enzyme Inhibitors, Malondialdehyde, MESH: Glycosylation End Products, Advanced, medicine.anatomical_structure, Losartan, Heterocyclic Compounds, 3-Ring, medicine.drug, medicine.medical_specialty, MESH: Rats, MESH: Diabetic Nephropathies, Biophysics, Internal medicine, medicine, Animals, Protease Inhibitors, Molecular Biology, MESH: Heterocyclic Compounds, 3-Ring, Angiotensin II receptor type 1, MESH: Angiotensin II Type 1 Receptor Blockers, Proteins, Angiotensin-converting enzyme, Cell Biology, MESH: Kidney, Rats, Rats, Zucker, Oxidative Stress, Endocrinology, biology.protein, Angiotensin II Type 1 Receptor Blockers
Abstract

International audience; Renin angiotensin system (RAS) worsens diabetic nephropathy (DN) by increasing oxidative stress. We compared the effect of three different RAS inhibitors: the angiotensin converting enzyme inhibitor Ramipril, the vasopeptidase inhibitor AVE7688 and the angiotensin receptor (AT1) antagonist Losartan on the formation of oxidative and carbonyl stress derived protein modifications in kidney from Zucker obese hyperglycemic rats (ZDFn Gm-fa/fa). Gas chromatography-mass spectrometry was used to measure representative markers of several protein oxidative pathways: direct oxidation [dinitrophenylhydrazine reactive carbonyls (DNP), glutamic (GSA), and aminoadipic (AASA) semialdehydes], mixed glyco- and lipoxidation [N(epsilon)-carboxyethyl-lysine (CEL) and N(epsilon)-(carboxymethyl)-lysine (CML)] and lipoxidation-[N(epsilon)-(malondialdehyde)-lysine-(MDAL)], as well as renal fatty acid composition. Urinary albumin (a marker of DN), DNP, GSA, and MDAL levels, were increased in all obese rats and were dose dependently decreased by AVE7688 whereas Ramipril and Losartan were less efficient. These results show that RAS inhibition improves DN at several levels, independently of its effects on blood pressure and glycemic control, via mechanisms depending of renal oxidative stress.

Published
2008

24. [Suppression of non-specific transcription by the proteasome in embryonic stem cells]

Author

Szutorisz, Henrietta, Dillon, Niall, Tora, Laszlo, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), and Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Immunoglobulin Light Chains, MESH: Protease Inhibitors, MESH: Proteasome Endopeptidase Complex, MESH: Transcription, Genetic, MESH: Membrane Glycoproteins, MESH: DNA, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Transcription Factors, MESH: Chromatin, MESH: B-Lymphocytes, MESH: Gene Expression Regulation, Developmental, MESH: Protein Binding, MESH: Animals, MESH: Models, Genetic, MESH: Embryonic Stem Cells, MESH: Mice
Published
2007

25. A library of novel hydroxamic acids targeting the metallo-protease family: design, parallel synthesis and screening

Author

Benoit Deprez, Terence Beghyn, Virginie Leroux, Rebecca Deprez-Poulain, Marion Flipo, Julie Charton, Charton, Julie, Biostructures et Decouverte de Medicament, Institut Pasteur de Lille, and 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, Droit et Santé

Subjects
MESH: Hydroxamic Acids, MESH: Enzyme Activation, Swine, medicine.medical_treatment, Clinical Biochemistry, MESH: Molecular Structure, Chemical biology, Drug Evaluation, Preclinical, Pharmaceutical Science, MESH: Drug Design, Hydroxamic Acids, Kidney, Biochemistry, Aminopeptidase, Aminopeptidases, MESH: Zinc, MESH: Aminopeptidases, MESH: Microsomes, Structure-Activity Relationship, MESH: Structure-Activity Relationship, Microsomes, Drug Discovery, medicine, Animals, Combinatorial Chemistry Techniques, Protease Inhibitors, MESH: Animals, Molecular Biology, MESH: Swine, chemistry.chemical_classification, Protease, MESH: Protease Inhibitors, Molecular Structure, Organic Chemistry, Stereoisomerism, MESH: Kidney, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, Combinatorial chemistry, MESH: Stereoisomerism, Enzyme Activation, [SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences, Zinc, Enzyme, chemistry, Drug Design, Molecular Medicine, MESH: Drug Evaluation, Preclinical, MESH: Combinatorial Chemistry Techniques
Abstract

International audience; We report here the design and parallel synthesis of 217 compounds based on a malonic-hydroxamic acid template. These compounds are obtained via a two-step solution-phase procedure. The set of diverse building-blocks used makes this strategy suitable for the search of inhibitors of various metallo-proteases and for the investigation of the biological role of new metallo-proteases. As a proof of concept, we screened this library on Neutral Aminopeptidase (APN; EC 3.4.11.2), the prototypal enzyme of the M1 family. Several submicromolar inhibitors were identified.

Published
2007

26. Prions: protein only or something more? Overview of potential prion cofactors

Author

Chiara Zurzolo, Carlo Fasano, Vincenza Campana, Physiologie neurovégétative - PNV (PNP), Université Paul Cézanne - Aix-Marseille 3-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Trafic membranaire et Pathogénèse, Institut Pasteur [Paris], Fasano, C, Campana, V, Zurzolo, Chiara, and Institut Pasteur [Paris] (IP)

Subjects
Protein Folding, PrPSc Proteins, Protein Conformation, animal diseases, Scrapie, Plasma protein binding, medicine.disease_cause, Transgenic, Prion Diseases, Mice, 0302 clinical medicine, Protein structure, MESH: Protein Conformation, Models, Protein Interaction Mapping, MESH: Cell-Free System, MESH: Animals, MESH: Brain Chemistry, MESH: Nerve Tissue Proteins, MESH: Lipid Metabolism, 0303 health sciences, Mutation, MESH: Protease Inhibitors, MESH: PrPSc Proteins, Intracellular Signaling Peptides and Proteins, General Medicine, Endocytosis, Biochemistry, MESH: Endocytosis, MESH: Scrapie, Protein folding, MESH: Molecular Chaperones, Protein Binding, Prions, MESH: Mice, Transgenic, Transgene, MESH: Protein Folding, MESH: Peptide Hydrolases, Mice, Transgenic, Nerve Tissue Proteins, Biology, Models, Biological, 03 medical and health sciences, Cellular and Molecular Neuroscience, MESH: Gene Expression Profiling, MESH: Prions, MESH: Intracellular Signaling Peptides and Proteins, medicine, Animals, Humans, MESH: Protein Binding, Protease Inhibitors, PrPC Proteins, MESH: PrPC Proteins, Gene, MESH: Mice, Protein Processing, 030304 developmental biology, Brain Chemistry, MESH: Humans, Cell-Free System, Gene Expression Profiling, MESH: Protein Interaction Mapping, Post-Translational, MESH: Models, Biological, MESH: Prion Diseases, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Lipid Metabolism, Biological, nervous system diseases, MESH: Solubility, Solubility, MESH: Protein Processing, Post-Translational, Protein Processing, Post-Translational, 030217 neurology & neurosurgery, Peptide Hydrolases, Molecular Chaperones
Abstract

Transmissible spongiform encephalopathies (TSEs) in humans and animals are attributed to protein-only infectious agents, called prions. Prions have been proposed to arise from the conformational conversion of the cellular protein PrP(C) into a misfolded form (e.g., PrP(Sc) for scrapie), which precipitates into aggregates and fibrils. It has been proposed that the conversion process is triggered by the interaction of the infectious form (PrP(Sc)) with the cellular form (PrP(C)) or might result from a mutation in the gene for PrP(C). However, until recently, all efforts to reproduce this process in vitro had failed, suggesting that host factors are necessary for prion replication. In this review we discuss recent findings such as the cellular factors that might be involved in the conformational conversion of prion proteins and the potential mechanisms by which they could operate.

Published
2006

27. The structural comparison of the bacterial PepX and human DPP-IV reveals sites for the design of inhibitors of PepX activity

Author

Pascal, Rigolet, Xu Guang, Xi, Stephane, Rety, Jean-François, Chich, Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), Unité de recherche Virologie et Immunologie Moléculaires (VIM (UR 0892)), Institut National de la Recherche Agronomique (INRA), Santé de la vigne et qualité du vin (SVQV), Institut National de la Recherche Agronomique (INRA)-Université Louis Pasteur - Strasbourg I, Université Louis Pasteur - Strasbourg I-Institut National de la Recherche Agronomique (INRA), and Unité de recherche Virologie et Immunologie Moléculaires (VIM)

Subjects
Models, Molecular, Adenosine Deaminase, Protein Conformation, Dipeptidyl Peptidase 4, [SDV]Life Sciences [q-bio], MESH: Antigens, CD26, MESH: Glycoproteins, MESH: Streptococcus, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH: Drug Design, MESH: Eukaryotic Cells, Substrate Specificity, MESH: Protein Conformation, Models, Adenosine Deaminase Inhibitors, Dipeptidyl Peptidases, Humans, Protease Inhibitors, Antigens, Dipeptidyl-Peptidases and Tripeptidyl-Peptidases, ComputingMilieux_MISCELLANEOUS, CD26, Glycoproteins, MESH: Adenosine Deaminase, Binding Sites, MESH: Protease Inhibitors, MESH: Humans, Molecular, Streptococcus, MESH: Dipeptidyl Peptidases, Lactococcus lactis, Eukaryotic Cells, MESH: Binding Sites, MESH: Lactococcus lactis, Drug Design, MESH: Substrate Specificity, MESH: Models, Molecular
Abstract

International audience; X-prolyl dipeptidyl aminopeptidases (X-PDAP) are enzymes catalysing the release of dipeptides from the amino termini of polypeptides containing a proline or an alanine at the penultimate position. Involved in various mammalian regulation processes, as well as in chronic human diseases, they have been proposed to play a role in pathogenicity for Streptococci. We compared the structure of X-PDAP from Lactococcus lactis (PepX) with its human counterpart DPP-IV. Despite very different overall folds, the residues most implicated for X-PDAP activity are conserved in the same positions and orientations in both enzymes, thus defining a structural signature for the X-PDAP specificity that crosses the species frontiers of evolution. Starting from this observation, we tested some inhibitors of DPP-IV on PepX activity, for which no specific inhibitor is known. We thus found that PepX was highly sensitive to valine-pyrrolidide with a KI of 9.3 microm, close to that reported in DPP-IV inhibition. We finally used the structure of PepX from L. lactis as a template for computer-based homology modeling of PepX from the pathogenic Streptococcus gordonii. Docking simulations of valine-pyrrolidide into the active site of PepX led to the identification of key residues for a rational drug design against PepX from Streptococci. These results could have applications in human health giving new perspectives to the struggle against pathogens.

Published
2005

28. Effect of doxycycline on sulfur mustard-induced respiratory lesions in guinea pigs

Author

Guignabert, Christophe, Taysse, Laurent, Calvet, Jean-Henri, Planus, Emmanuelle, Delamanche, Séraphin, Galiacy, Stéphane, d'Ortho, Marie-Pia, Guignabert, Christophe, Physiopathologie et Thérapeutiques Respiratoires, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10, Centre d'Etudes du Bouchet, Délégation Générale pour l'Armement, Service de physiologie, explorations fonctionnelles [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), and This work was funded by the DGA and the Institut National de la Sante et de la Recherche Medicale (France).

Subjects
MESH: Inflammation, [SDV.MHEP.PHY] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], [CHIM.THER] Chemical Sciences/Medicinal Chemistry, MESH: Chemical Warfare Agents, protease inhibitors, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, MESH: Guinea Pigs, [SDV.MHEP.CSC]Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, MESH: Anti-Bacterial Agents, vesicant agents, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], MESH: Animals, MESH: Gelatinases, MESH: Doxycycline, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, MESH: Respiratory System, MESH: Protease Inhibitors, doxycycline, blistering agents, MESH: Male, [SDV.MHEP.CSC] Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, [SDV.TOX] Life Sciences [q-bio]/Toxicology, MESH: Mustard Gas, MESH: Tissue Inhibitor of Metalloproteinase-2, MESH: Tissue Inhibitor of Metalloproteinase-1, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, MESH: Epithelial Cells, [SDV.TOX]Life Sciences [q-bio]/Toxicology, [SDV.MHEP.PSR] Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, proteases, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie
Abstract

9 pages; International audience; Respiratory tract lesions induced by the chemical warfare agent sulfur mustard (SM) are characterized by epithelial damages associated with inflammatory cell infiltration. Here we evaluated the imbalance between gelatinase and tissue inhibitors of metalloproteinases (TIMPs), and we tested pretreatment with the protease inhibitor doxycycline. Guinea pigs were intoxicated intratracheally with SM and evaluated 24 h after exposure. Matrix metalloproteinase (MMP) gelatinase activity of bronchial lavage (BL) fluid from SM-exposed guinea pigs was high compared with controls, as shown by both zymography and biotinylated substrate degradation, whereas TIMP-1 and -2 levels by immunoblotting were similar. Extensive areas of lysis were evidenced by in situ zymography, indicating imbalance between gelatinases and inhibitors towards net proteolytic activity. Doxycycline pretreatment resulted in 1) decreased gelatinase activity (zymography, free gelatinase activity assay, and in situ zymography); 2) decreased inflammation (BL fluid cellularity and protein level); and 3) dramatic decrease in histological epithelial lesions. Our results suggest inadequate levels of TIMP to counteract increased gelatinase activity and further support a role for MMP gelatinases in SM-induced respiratory lesions. They also suggest that doxycycline may hold promise as a therapeutic tool.

Published
2005

29. Synthesis of small molecule CDC25 phosphatases inhibitors

Author

Bernard Ducommun, Gregoire Prevost, Marie-Christine Brezak, Marie-Odile Contour-Galcera, Olivier Lavergne, Institut Henri Beaufour (IPSEN), IPSEN-BEAUFOUR, Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération (LBCMCP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cdc25, Clinical Biochemistry, Nitro compound, Pharmaceutical Science, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biochemistry, Chemical synthesis, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, MESH: cdc25 Phosphatases, Drug Discovery, Humans, cdc25 Phosphatases, Protease Inhibitors, Molecular Biology, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, MESH: Humans, MESH: Protease Inhibitors, biology, Chemistry, Organic Chemistry, Biological activity, Small molecule, 3. Good health, enzymes and coenzymes (carbohydrates), Enzyme, Enzyme inhibitor, 030220 oncology & carcinogenesis, biology.protein, Molecular Medicine, biological phenomena, cell phenomena, and immunity, Lead compound
Abstract

A targeted library of small molecules has been prepared to optimize the biological activity of BN82002, our initial lead compound, recently described as an original inhibitor of CDC25 phosphatases. Some of these compounds inhibit CDC25 in the micromolar range and therefore reinforce the interest of CDC25 as an anticancer target.

Published
2004

30. When hypoxia signalling meets the ubiquitin-proteasomal pathway, new targets for cancer therapy

Author

Jacques Pouysségur, Christiane Brahimi-Horn, Institut de signalisation, biologie du développement et cancer (ISBDC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA)

Subjects
MAPK/ERK pathway, MESH: Signal Transduction, Proteasome Endopeptidase Complex, MESH: Ubiquitin, MESH: Anoxia, [SDV.CAN]Life Sciences [q-bio]/Cancer, 03 medical and health sciences, 0302 clinical medicine, Ubiquitin, Neoplasms, Humans, MESH: Neoplasms, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Protease Inhibitors, Hypoxia, 030304 developmental biology, G alpha subunit, 0303 health sciences, MESH: Humans, MESH: Protease Inhibitors, biology, MESH: Proteasome Endopeptidase Complex, Hematology, MESH: Transcription Factors, Hedgehog signaling pathway, 3. Good health, Cell biology, Oncology, Proteasome, Biochemistry, Ubiquitin-Proteasomal Pathway, 030220 oncology & carcinogenesis, MESH: Protein Processing, Post-Translational, biology.protein, Mdm2, Signal transduction, Protein Processing, Post-Translational, Signal Transduction, Transcription Factors
Abstract

International audience; The ubiquitin-proteasomal pathway of degradation of proteins is activated or repressed in response to a number of environmental stresses and thereby plays an essential role in cell function and survival. Hypoxic stress, resulting from a decrease in the concentration of oxygen in tissues, is encountered in both physiological and pathological situations, in particular in cancer. The transcriptional complex hypoxia-inducible factor (HIF) is the key player in the signalling pathway that controls the hypoxic response of mammalian cells. Under hypoxic conditions it transactivates an impressive number of genes involved in a multitude of cellular functions. Tight regulation of this response in part involves the ubiquitin-proteasomal system where oxygen-dependent prolyl-4-hydroxylation of the alpha subunit of HIF triggers a cascade of events that leads to its degradation by the 26S proteasome. Inhibition of the proteasome in conjunction with topoisomerase inhibition has shown some promise in the treatment of experimental cancer. Such treatment may impact on the hypoxic adaptation of tumour cells.

Published
2004

31. In Vivo Inhibition of Osteoblastic Metalloproteinases Leads to Increased Trabecular Bone Mass

Author

Catherine Terraz, Valérie Geoffroy, Caroline Marty-Morieux, Marie Christine de Vernejoul, Phillippe Clement-Lacroix, Sophie Roux, Jerome Rossert, Nathalie Le Goupil, Monique Frain, Os et articulations, Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Génétique moléculaire du développement, Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (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)-Centre National de la Recherche Scientifique (CNRS)-IFR36-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Male, MESH: Bone Resorption, Endocrinology, Diabetes and Metabolism, Parathyroid hormone, Bone remodeling, Mice, 0302 clinical medicine, MESH: Alkaline Phosphatase, Bone Density, Orthopedics and Sports Medicine, MESH: Animals, Promoter Regions, Genetic, MESH: Radiography, MESH: Bone Density, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Cells, Cultured, 0303 health sciences, MESH: Protease Inhibitors, Chemistry, Osteoblast, MESH: Bone and Bones, Resorption, medicine.anatomical_structure, Phenotype, 030220 oncology & carcinogenesis, Osteocyte, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Metalloproteases, MESH: Cells, Cultured, medicine.medical_specialty, MESH: Mice, Transgenic, Mice, Transgenic, [SDV.CAN]Life Sciences [q-bio]/Cancer, Bone healing, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH: Phenotype, Bone resorption, Bone and Bones, 03 medical and health sciences, Osteoclast, MESH: Mice, Inbred C57BL, Internal medicine, MESH: Promoter Regions, Genetic, medicine, Animals, Protease Inhibitors, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Bone Resorption, MESH: Mice, 030304 developmental biology, MESH: Osteoblasts, [SDV.GEN]Life Sciences [q-bio]/Genetics, Osteoblasts, Tissue Inhibitor of Metalloproteinase-1, Alkaline Phosphatase, MESH: Male, Mice, Inbred C57BL, Radiography, Endocrinology, MESH: Tissue Inhibitor of Metalloproteinase-1, Metalloproteases, MESH: Female
Abstract

Mice specifically overexpressing TIMP-1 in osteoblasts have been generated to investigate the role of MMPs in bone in vivo. These mice displayed increased trabecular bone volume and decreased bone turnover. This model provides evidence of the role played by the MMPs in bone remodeling and balance. Introduction: Although it has been suggested that the matrix metalloproteinases (MMPs) may play a role in initiating the bone resorption process in vitro, there is no evidence that they play any role in in vivo bone maintenance. Materials and Methods: We used an artificial promoter specifically driving cells of the osteoblastic lineage to overexpress the tissue inhibitor of MMPs (TIMP-1) cDNA in mice. Densitometric analysis, using DXA and pQCT, and static and dynamic histomorphometry were used to evaluate the bone phenotype both in male and female transgenic mice. We evaluated osteoblastic differentiation using a primary osteoblast culture and osteoclast activity using an ex vivo organ culture. Results and Conclusion: We showed that at 1 and 2.5 months of age, only the female mice exhibited a bone phenotype. These mice displayed specific increases in the BMD and bone volume of trabecular bone. This increase was accompanied by decreased trabecular separation, suggesting a decrease in bone resorption. Using an ex vivo resorption assay, we demonstrated that parathyroid hormone (PTH)-stimulated bone resorption was reduced in these mice. Evaluation of the bone histomorphometric dynamic parameters showed that the mineralizing surfaces and bone formation rate were both reduced. There was no change in the mineralization lag time or number of osteocyte lacunae. Using primary osteoblast culture and molecular analysis, we showed that the differentiation and function of osteoblasts from transgenic mice were normal, but that the ex vivo formation of mineralized nodules was delayed. This model is the first to show that in vivo MMPs play a role in bone remodeling and bone balance. Moreover, our data suggest that MMP activity could be involved in the hormonal regulation of bone resorption by osteoblasts. J Bone Miner Res 2004;19:811- 822. Published online on January 19, 2004; doi: 10.1359/JBMR.040119

Published
2004

32. Sialorphin, a natural inhibitor of rat membrane-bound neutral endopeptidase that displays analgesic activity

Author

Catherine Rougeot, Thierry Blisnick, Anne Gaëlle Rigault, Christophe Dugave, François Rougeon, Didier Desor, Véronique Hermitte, Michaël Messaoudi, Génétique et Biochimie du Développement, Institut Pasteur [Paris], ETAP-Applied Ethology, Partenaires INRAE, Biologie des Interactions Hôte-Parasite, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Département d'Ingenierie et d'Etudes des Protéines (DIEP), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Henri Poincaré - Nancy 1 (UHP), Institut Pasteur [Paris] (IP), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, MESH: Protein Precursors, MESH: Analgesics, MESH: Substance P, Receptors, Opioid, mu, Endogeny, Substance P, MESH: Amino Acid Sequence, MESH: Pain Measurement, Kidney, MESH: Formaldehyde, MESH: Spinal Cord, chemistry.chemical_compound, 0302 clinical medicine, Receptors, Opioid, delta, MESH: Receptors, Opioid, delta, MESH: Animals, MESH: Submandibular Gland, Receptor, Neprilysin, Pain Measurement, Endogenous opioid, Analgesics, 0303 health sciences, Multidisciplinary, MESH: Protease Inhibitors, Chemistry, MESH: Neprilysin, Glycopeptides, Prostate, MESH: Salivary Proteins and Peptides, Biological Sciences, MESH: Enkephalin, Methionine, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, Naltrexone, 3. Good health, Spinal Cord, MESH: Naltrexone, MESH: Pain, MESH: Membrane Proteins, medicine.drug, Thiorphan, medicine.medical_specialty, MESH: Rats, Enkephalin, Methionine, Molecular Sequence Data, Submandibular Gland, Pain, MESH: Receptors, Opioid, mu, MESH: Prostate, 03 medical and health sciences, Leucine, In vivo, Formaldehyde, Internal medicine, medicine, Animals, Protease Inhibitors, Amino Acid Sequence, Protein Precursors, Rats, Wistar, Salivary Proteins and Peptides, 030304 developmental biology, MESH: Glycopeptides, MESH: Molecular Sequence Data, Opiorphin, Membrane Proteins, MESH: Thiorphan, MESH: Rats, Wistar, MESH: Kidney, In vitro, MESH: Male, Rats, Endocrinology, MESH: Leucine, MESH: Wounds and Injuries, Wounds and Injuries, 030217 neurology & neurosurgery
Abstract

Sialorphin is an exocrine and endocrine signaling mediator, which has been identified by a genomic approach. It is synthesized predominantly in the submandibular gland and prostate of adult rats in response to androgen steroids and is released locally and systemically in response to stress. We now demonstrate that the cell surface molecule to which sialorphin binds in vivo in the rat kidney is the membrane-anchored neutral endopeptidase (neprilysin; NEP, EC 3.4.24.11). NEP plays an important role in nervous and peripheral tissues, as it turns off several peptide-signaling events at the cell surface. We show that sialorphin prevents spinal and renal NEP from breaking down its two physiologically relevant substrates, substance P and Met-enkephalin in vitro . Sialorphin inhibited the breakdown of substance P with an IC 50 of 0.4–1 μM and behaved as a competitive inhibitor. In vivo , i.v. sialorphin elicited potent antinociceptive responses in two behavioral rat models of injury-induced acute and tonic pain, the pin-pain test and formalin test. The analgesia induced by 100–200 μg/kg doses of sialorphin required the activation of μ- and δ-opioid receptors, consistent with the involvement of endogenous opioid receptors in enkephalinergic transmission. We conclude that sialorphin protects endogenous enkephalins released after nociceptive stimuli by inhibiting NEP in vivo . Sialorphin is a natural systemically active regulator of NEP activity. Furthermore, our study provides evidence that it is a physiological modulator of pain perception after injury and might be the progenitor of a new class of therapeutic molecules.

Published
2003

33. Proteasome inhibitors as therapeutic agents: current and future strategies

Author

Claude Prigent, M. Baudy Floc'h, Jean-Guy Delcros, Yannick Arlot-Bonnemains, Groupe de Recherche en Thérapeutique Anticancéreuse, Université de Rennes (UR), Génétique et Développement, Université de Rennes (UR)-IFR97-Centre National de la Recherche Scientifique (CNRS), Synthèse et électrosynthèse organiques (SESO), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Prigent, Claude, Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-IFR97-Centre National de la Recherche Scientifique (CNRS), and Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Proteases, Programmed cell death, Proteasome Endopeptidase Complex, Cell, Antineoplastic Agents, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Protein degradation, Biology, MESH: Multienzyme Complexes, 01 natural sciences, Biochemistry, MESH: Neurodegenerative Diseases, 03 medical and health sciences, 0302 clinical medicine, Ubiquitin, Multienzyme Complexes, Neoplasms, Drug Discovery, medicine, Animals, Humans, MESH: Ubiquitins, Protease Inhibitors, MESH: Animals, MESH: Neoplasms, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Ubiquitins, Pharmacology, MESH: Humans, MESH: Protease Inhibitors, 010405 organic chemistry, MESH: Proteasome Endopeptidase Complex, Organic Chemistry, Neurodegenerative Diseases, Cell cycle, 0104 chemical sciences, Cysteine Endopeptidases, medicine.anatomical_structure, Proteasome, 030220 oncology & carcinogenesis, biology.protein, Molecular Medicine, MESH: Antineoplastic Agents, Intracellular, MESH: Cysteine Endopeptidases
Abstract

International audience; In cells, protein degradation is a key pathway for the destruction of abnormal or damaged proteins as well as for the elimination of proteins whose presence is no longer required. Among the various cell proteases, the proteasome, a multicatalytic macromolecular complex, is specifically required for the degradation of ubiquitinated proteins. In normal cells, the proteasome ensures the elimination of numerous proteins that play critical roles in cell functions throughout the cell cycle. Defects in the activity of this proteolytic machinery can lead to the disorders of cell function that is believed to be the root cause of certain diseases. Indeed, many proteins involved in the control of cell cycle transitions are readily destroyed by the proteasome once their tasks have been accomplished. Moreover, because proteasome inhibitors can provoke cell death, it has been suggested that proteasomes must be continually degrading certain apoptotic factors. For these reasons, proteasome inhibition has become a new and potentially significant strategy for the drug development in cancer treatment. The proteasome possesses three major peptidase activities that can individually be targeted by drugs. Different classes of proteasome inhibitors are reviewed here. In addition, we present new pseudopeptides with the enriched nitrogen backbones bearing a side chain and a modified C-terminal position that inhibit proteasome activity.

Published
2003

34. General function of N-terminal propeptide on assisting protein folding and inhibiting catalytic activity based on observations with a chimeric thermolysin-like protease

Author

Motomitsu Kitaoka, Satoru Nirasawa, Cynthia Marie-Claire, Kiyoshi Hayashi, Bing Tang, Enzyme Laboratory, Biological Function Division, National Food Research Institute, Pharmacochimie moléculaire et structurale, 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)-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 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) - 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) - 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)-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)-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)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Marie-Claire, Cynthia

Subjects
Protein Folding, MESH: Aeromonas, medicine.medical_treatment, Biochemistry, MESH: Enzyme Precursors, MESH : Bacterial Proteins, MESH : Catalysis, MESH: Endopeptidases, MESH: Bacterial Proteins, chemistry.chemical_classification, 0303 health sciences, Metalloproteinase, Enzyme Precursors, MESH: Protease Inhibitors, biology, MESH : Aeromonas, MESH : Enzyme Precursors, Metalloendopeptidases, MESH : Endopeptidases, MESH : Metalloendopeptidases, Protein folding, Aeromonas, MESH : Protein Folding, Proteases, MESH : Protease Inhibitors, Vibriolysin, MESH : Recombinant Fusion Proteins, Recombinant Fusion Proteins, MESH: Protein Folding, Biophysics, MESH: Metalloendopeptidases, Catalysis, 03 medical and health sciences, Bacterial Proteins, Endopeptidases, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, medicine, MESH: Recombinant Fusion Proteins, Protease Inhibitors, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Protein precursor, Molecular Biology, MESH : Protein Processing, Post-Translational, 030304 developmental biology, Protease, 030306 microbiology, Cell Biology, MESH: Catalysis, Enzyme, chemistry, Chaperone (protein), MESH: Protein Processing, Post-Translational, biology.protein, Protein Processing, Post-Translational
Abstract

Pro-aminopeptidase processing protease (PA protease) is a thermolysin-like metalloprotease produced by Aeromonas caviae T-64. The N-terminal propeptide acts as an intramolecular chaperone to assist the folding of PA protease and shows inhibitory activity toward its cognate mature enzyme. Moreover, the N-terminal propeptide strongly inhibits the autoprocessing of the C-terminal propeptide by forming a complex with the folded intermediate pro-PA protease containing the C-terminal propeptide (MC). In order to investigate the structural determinants within the N-terminal propeptide that play a role in the folding, processing, and enzyme inhibition of PA protease, we constructed a chimeric pro-PA protease by replacing the N-terminal propeptide with that of vibriolysin, a homologue of PA protease. Our results indicated that, although the N-terminal propeptide of vibriolysin shares only 36% identity with that of PA protease, it assists the refolding of MC, inhibits the folded MC to process its C-terminal propeptide, and shows a stronger inhibitory activity toward the mature PA protease than that of PA protease. These results suggest that the N-terminal propeptide domains in these thermolysin-like proteases may have similar functions, in spite of their primary sequence diversity. In addition, the conserved regions in the N-terminal propeptides of PA protease and vibriolysin may be essential for the functions of the N-terminal propeptide.

Published
2003

35. Impact of protease inhibitors on intrahepatic hepatitis C virus viral load

Author

Michel Dupon, Marie-Edith Lafon, Jean-Marie Ragnaud, Pascale Trimoulet, Elisabeth Legrand, Didier Neau, Brigitte Le Bail, Maria Winnock, Evelyne Schvoerer, Hervé Fleury, Service de virologie et d'immunologie biologique, CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, Imagerie moléculaire et fonctionnelle: de la physiologie à la thérapie, Université Bordeaux Segalen - Bordeaux 2-IFR8-Centre National de la Recherche Scientifique (CNRS), Ecogéochimie et Fonctionnement des Ecosystèmes Benthiques (EFEB), Adaptation et diversité en milieu marin (AD2M), 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)-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), CHU Bordeaux [Bordeaux], CHU Pellegrin, Laboratory for lasers energetics - LLE (New-York, USA), and University of Rochester [USA]

Subjects
Male, Hepacivirus, HIV Infections, medicine.disease_cause, MESH: Antiretroviral Therapy, Highly Active, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Antiretroviral Therapy, Highly Active, Immunology and Allergy, MESH: Hepacivirus, 030212 general & internal medicine, Sida, MESH: Middle Aged, MESH: Protease Inhibitors, biology, virus diseases, MESH: HIV Infections, Middle Aged, 3. Good health, MESH: Hepatitis C, Chronic, Infectious Diseases, Liver, MESH: RNA, Viral, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, RNA, Viral, Female, 030211 gastroenterology & hepatology, Viral disease, Viral load, Adult, Hepatitis C virus, Immunology, Virus, 03 medical and health sciences, Flaviviridae, medicine, Humans, Protease Inhibitors, Protease inhibitor (pharmacology), MESH: Humans, business.industry, MESH: Adult, Hepatitis C, Chronic, biology.organism_classification, Virology, digestive system diseases, MESH: Male, business, MESH: Female, MESH: Liver
Abstract

International audience; During chronic hepatitis C, hepatitis C virus (HCV) load in plasma was shown to be higher in HIV-co-infected than in immunocompetent patients [1]. The reason for this increased HCV replication is not known. It may be as a result of HIV-induced immune deficiency [2], although some authors did not find any correlation with the CD4 cell count [3]. A direct interaction between HCV and HIV was also hypothesized [4]. Protease inhibitors (PI) used in highly active antiretroviral therapy (HAART) have no HCV reduction effect during the first months of treatment [5-8]. However, a decrease in HCV plasma load was recently described in patients treated with HAART for a year [9,10]. We therefore investigated the potential impact of HAART on intrahepatic HCV load.

Published
2001

36. Differences in transition state stabilization between thermolysin (EC 3.4.24.27) and neprilysin (EC 3.4.24.11)

Author

Marie-Claude Fournie-Zaluski, Cynthia Marie-Claire, Emmanuel Ruffet, Gilles Tiraboschi, Pharmacochimie moléculaire et structurale, 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)-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)-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 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)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Marie-Claire, Cynthia, Institut des sciences du Médicament -Toxicologie - Chimie - Environnement ( IFR71 ), and 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 ) -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 ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
Models, Molecular, MESH: Enzyme Stability, Molecular model, Protein Conformation, MESH : Aspartic Acid, MESH: Catalytic Domain, MESH: Amino Acid Sequence, Biochemistry, MESH: Aspartic Acid, MESH: Recombinant Proteins, chemistry.chemical_compound, 0302 clinical medicine, Protein structure, MESH: Protein Conformation, Structural Biology, Catalytic Domain, Enzyme Stability, Aspartic acid, Site-directed mutagenesis, Neprilysin, MESH : Protein Conformation, MESH : Neprilysin, 0303 health sciences, MESH: Protease Inhibitors, MESH: Kinetics, Chemistry, MESH: Neprilysin, MESH : Amino Acid Sequence, Glycopeptides, MESH : Catalytic Domain, MESH : Bacillus subtilis, Recombinant Proteins, MESH : Mutagenesis, Site-Directed, MESH: Mutagenesis, Site-Directed, medicine.symptom, MESH : Kinetics, MESH : Th, MESH: Models, Molecular, Bacillus subtilis, Thiorphan, MESH : Glycopeptides, MESH : Protease Inhibitors, MESH : Recombinant Proteins, MESH : Models, Molecular, Biophysics, Thermolysin, Molecular modeling, MESH: Thermolysin, 03 medical and health sciences, Genetics, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Protease Inhibitors, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Molecular Biology, MESH : Thermolysin, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Th, 030304 developmental biology, MESH: Glycopeptides, Aspartic Acid, MESH : Enzyme Stability, Phosphoramidon, fungi, Cell Biology, MESH: Bacillus subtilis, Kinetics, Mechanism of action, Mutagenesis, Site-Directed, Tyrosine, 030217 neurology & neurosurgery
Abstract

Important homologies in the topology of the catalytic site and the mechanism of action of thermolysin and neprilysin have been evidenced by site-directed mutagenesis. The determination of differences in transition state stabilization between these peptidases could facilitate the design of specific inhibitors. Thus, two residues of thermolysin which could be directly (Tyr157) or indirectly (Asp226) involved in the stabilization of the transition state and their putative counterparts in neprilysin (Tyr625 and Asp709) have been mutated. The results show that Tyr157 is important for thermolysin activity while Tyr625 has no functional role in neprilysin. Conversely, the mutation of Asp226 induced a slight perturbation of thermolysin activity while Asp709 in neprilysin seems crucial in neprilysin catalysis. Taken together these data seem to indicate differences in the transition state mode of stabilization in the two peptidases.

Published
1998

37. Characterization of Glu350 as a critical residue involved in the N-terminal amine binding site of aminopeptidase N (EC 3.4.11.2): insights into its mechanism of action

Author

Marie-Claude Fournie-Zaluski, Bernard P. Roques, Ann Beaumont, Emmanuel Ruffet, Nathalie Luciani, Cynthia Marie-Claire, Pharmacochimie moléculaire et structurale, 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)-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)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Marie-Claire, Cynthia, 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)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and 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) - 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) - Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS)

Subjects
Swine, MESH: Amino Acid Sequence, Biochemistry, 0302 clinical medicine, MESH: Animals, chemistry.chemical_classification, 0303 health sciences, MESH: Protease Inhibitors, biology, MESH: Antigens, CD13, MESH: Models, Chemical, Amine binding, MESH: Glutamic Acid, Recombinant Proteins, Amino acid, MESH: COS Cells, MESH: Mutagenesis, Site-Directed, 030220 oncology & carcinogenesis, COS Cells, medicine.symptom, Protein Binding, MESH: Mutation, Stereochemistry, MESH: Peptide Hydrolases, Molecular Sequence Data, chemistry.chemical_element, Glutamic Acid, Zinc, CD13 Antigens, 03 medical and health sciences, Residue (chemistry), Exopeptidases, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Protein Binding, Animals, MESH: Exopeptidases, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Protease Inhibitors, Amino Acid Sequence, Binding site, 030304 developmental biology, MESH: Molecular Sequence Data, Binding Sites, Sequence Homology, Amino Acid, Exopeptidase, Enzyme, Mechanism of action, chemistry, MESH: Binding Sites, Models, Chemical, MESH: Recombinant Pr, Mutation, biology.protein, Mutagenesis, Site-Directed, Peptide Hydrolases
Abstract

The molecular components ensuring the strict exopeptidase action of aminopeptidase N (APN) and related zinc aminopeptidases of the M1 family have not yet been clearly established. The specific recognition of the N-terminal amino acid of the substrates by the enzymes has been proposed to involve either the complexation of the free amino group by the catalytic zinc ion or an interaction with an anionic binding site, which could be constituted by an aspartate or glutamate residue. To investigate the existence of such an ionic binding site, site-directed mutagenesis experiments have been performed on acidic residues of pig APN. Given that aminopeptidases of the M1 family are likely to have a common mechanism of action, only strictly conserved residues were mutated. As compared to the wild-type enzyme, the mutation D220E led only to slight modifications in the kinetic parameters of the enzyme and in the Ki values of various inhibitors, indicating that this residue is not critically involved in the hydrolytic mechanism. In contrast, the mutations E350Q and E350D induced a large decrease in enzyme activity, essentially due to modifications in kcat, whereas the E350A mutation led to an almost completely inactive enzyme. Moreover, among the inhibitors tested, only those acting as transition state analogs showed significant increases in their Ki values. These data are in favor of E350 belonging to the "anionic binding site" in APN. A mechanism of action, derived from that of thermolysin, is proposed for these aminopeptidases, which explains the importance of E350 in transition state formation, rather than in the Michaelis complex.

Published
1998

38. Plasmodium chabaudi p68 serine protease activity required for merozoite entry into mouse erythrocytes

Author

L. Pereira da Silva, Gordon Langsley, Thierry Blisnick, Thierry Rabilloud, Catherine Braun Breton, Hélène Jouin, Jean-Christophe Barale, Parasitologie Expérimentale, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Biochimie et Génétique, and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)

Subjects
Erythrocytes, 030231 tropical medicine, MESH: Erythrocyte Membrane, Substrate Specificity, Plasmodium chabaudi, 03 medical and health sciences, Mice, 0302 clinical medicine, Gentamicin protection assay, Anion Exchange Protein 1, Erythrocyte, medicine, Animals, MESH: Animals, [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology, Protease Inhibitors, MESH: Serine Endopeptidases, Band 3, MESH: Mice, 030304 developmental biology, Serine protease, chemistry.chemical_classification, 0303 health sciences, Multidisciplinary, MESH: Protease Inhibitors, biology, MESH: Erythrocytes, MESH: Molecular Weight, Erythrocyte Membrane, Serine Endopeptidases, biology.organism_classification, Molecular biology, In vitro, 3. Good health, MESH: Plasmodium chabaudi, Molecular Weight, Red blood cell, medicine.anatomical_structure, Enzyme, Biochemistry, chemistry, Enzyme inhibitor, biology.protein, MESH: Substrate Specificity, MESH: Anion Exchange Protein 1, Erythrocyte, Research Article
Abstract

International audience; To define the role of malaria parasite enzymes during the process of erythrocyte invasion, we have developed an in vitro serum-free invasion assay of mouse erythrocytes by purified Plasmodium chabaudi merozoites. The sensitivity of a merozoite-specific serine protease (p68) to various inhibitors and the effect of these inhibitors on invasion indicate a crucial role for p68. The substrate specificity of the purified enzyme has been partially defined using fluorogenic peptides. Consistent with this, in vitro incubation of mouse erythrocytes with the merozoite enzyme led to the cleavage of band 3 protein. The possible implication of erythrocyte band 3 truncation for the successful entry of the merozoite into the erythrocyte is discussed.

Published
1992

39. A Family of Diverse Kunitz Inhibitors from Echinococcus granulosus Potentially Involved in Host-Parasite Cross-Talk

Author

John Parkinson, Martín Fló, Cecilia Fernández, Silvia González, Rosario Durán, Rick M. Maizels, Beatriz Alvarez, Martín Graña, Gualberto González-Sapienza, Gustavo Salinas, Mariana Margenat, Cátedra de Inmunología, Facultad de Quimica, Laboratorio de Enzimología, Facultad de Ciencias, Institut Pasteur de Montevideo, Réseau International des Instituts Pasteur (RIIP), Hospital for Sick Children, Institute of Immunology and Infection Research, and University of Edinburgh

Subjects
MESH: Hydrogen-Ion Concentration, MESH: Sequence Homology, Amino Acid, MESH: Serine Proteases, lcsh:Medicine, MESH: Amino Acid Sequence, MESH: Echinococcus granulosus, MESH: Dose-Response Relationship, Drug, MESH: Dogs, Serine, MESH: Protein Structure, Tertiary, Parasite hosting, MESH: Animals, lcsh:Science, Echinococcus granulosus, Biochemistry/Biomacromolecule-Ligand Interactions, Genes, Helminth, 0303 health sciences, MESH: Protease Inhibitors, Multidisciplinary, biology, Genetics and Genomics/Functional Genomics, 030302 biochemistry & molecular biology, Hydrogen-Ion Concentration, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Biochemistry/Molecular Evolution, Biochemistry, MESH: Pepsin A, Genetics and Genomics/Gene Discovery, Research Article, Proteases, Molecular Sequence Data, Biochemistry/Biocatalysis, MESH: Host-Parasite Interactions, Host-Parasite Interactions, MESH: Gene Expression Profiling, 03 medical and health sciences, Dogs, Complementary DNA, parasitic diseases, Animals, Protease Inhibitors, Amino Acid Sequence, Gene, Infectious Diseases/Helminth Infections, 030304 developmental biology, MESH: Molecular Sequence Data, Dose-Response Relationship, Drug, Sequence Homology, Amino Acid, Host (biology), Gene Expression Profiling, lcsh:R, biology.organism_classification, Pepsin A, Protein Structure, Tertiary, Gene expression profiling, lcsh:Q, Serine Proteases, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM], MESH: Genes, Helminth
Abstract

International audience; The cestode Echinococcus granulosus, the agent of hydatidosis/echinococcosis, is remarkably well adapted to its definitive host. However, the molecular mechanisms underlying the successful establishment of larval worms (protoscoleces) in the dog duodenum are unknown. With the aim of identifying molecules participating in the E. granulosus-dog cross-talk, we surveyed the transcriptomes of protoscoleces and protoscoleces treated with pepsin at pH 2. This analysis identified a multigene family of secreted monodomain Kunitz proteins associated mostly with pepsin/H(+)-treated worms, suggesting that they play a role at the onset of infection. We present the relevant molecular features of eight members of the E. granulosus Kunitz family (EgKU-1 - EgKU-8). Although diverse, the family includes three pairs of close paralogs (EgKU-1/EgKU-4; EgKU-3/EgKU-8; EgKU-6/EgKU-7), which would be the products of recent gene duplications. In addition, we describe the purification of EgKU-1 and EgKU-8 from larval worms, and provide data indicating that some members of the family (notably, EgKU-3 and EgKU-8) are secreted by protoscoleces. Detailed kinetic studies with native EgKU-1 and EgKU-8 highlighted their functional diversity. Like most monodomain Kunitz proteins, EgKU-8 behaved as a slow, tight-binding inhibitor of serine proteases, with global inhibition constants (K(I) (*)) versus trypsins in the picomolar range. In sharp contrast, EgKU-1 did not inhibit any of the assayed peptidases. Interestingly, molecular modeling revealed structural elements associated with activity in Kunitz cation-channel blockers. We propose that this family of inhibitors has the potential to act at the E. granulosus-dog interface and interfere with host physiological processes at the initial stages of infection.

Published
2009

40. Evidence for pyroglutamyl peptidase I and prolyl endopeptidase activities in the rat insulinoma cell line RINm 5F: lack of relationship with TRH metabolism

Author

P. Salers, Anne Dutour, L'Houcine Ouafik, Pierre Giraud, J. Y. Maltese, Charles Oliver, Ouafik, L'Houcine, INSERM U 297 - Laboratoire de Neuroendocrinologie Expérimentale, and Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
endocrine system diseases, Clinical Biochemistry, Thyrotropin-releasing hormone, MESH: Amino Acid Sequence, Substrate Specificity, MESH: Pyroglutamyl-Peptidase I, MESH: Animals, MESH: Serine Endopeptidases, MESH: Endopeptidases, Thyrotropin-Releasing Hormone, chemistry.chemical_classification, MESH: Protease Inhibitors, MESH: Kinetics, MESH: Molecular Weight, Serine Endopeptidases, General Medicine, Adenoma, Islet Cell, Biochemistry, Enzyme inhibitor, Cell fractionation, Prolyl Oligopeptidases, MESH: Diazomethane, hormones, hormone substitutes, and hormone antagonists, Subcellular Fractions, medicine.drug, endocrine system, MESH: Rats, Molecular Sequence Data, Radioimmunoassay, MESH: Adenoma, Islet Cell, Pyroglutamyl-Peptidase I, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, Peptide hormone, Cell Line, MESH: Radioimmunoassay, [SDV.CAN] Life Sciences [q-bio]/Cancer, Prolyl endopeptidase, MESH: Thyrotropin-Releasing Hormone, Endopeptidases, medicine, Animals, Protease Inhibitors, Amino Acid Sequence, Pyroglutamyl-peptidase I, Molecular Biology, MESH: Molecular Sequence Data, Rat Insulinoma, Cell Biology, MESH: Cell Line, Rats, Molecular Weight, Kinetics, Enzyme, Diazomethane, chemistry, MESH: Subcellular Fractions, biology.protein, MESH: Substrate Specificity
Abstract

International audience; Thyrotropin-Releasing hormone (TRH)-degrading pyroglutamyl peptidase I (PGP I) and prolylendopeptidase (PE) activities have been demonstrated in rat insulinoma RINm 5F cell line. These two enzymes catalyze the conversion of TRH to Histydyl-Proline-Diketopiperazine and to acid TRH respectively. After cell fractionation, we found all the PGP I and PE activities in the cytosolic fraction. The membrane-bound PGP II activity is not detectable in the RINm 5F cells. Further investigations on these two cytosolic enzymes show that pyroglutamyl- and proline-containing peptides are inhibitors of each TRH-degrading enzyme. Gel filtration chromatography on Sephadex G100 shows that PGP I and PE activity have an apparent molecular mass of about 18 kDa and 57 kDa, respectively. Kinetic analysis with TRH as substrate, gives a Km of 44 microM and 235 microM, and a Vmax of 1.49 and 8.80 pmol/min/micrograms protein for PGP I and PE, respectively. Immunoreactive TRH, His-Pro-Diketopiperazine and acid TRH levels in the cell line extracts are 2.2 +/- 0.9, 22.5 +/- 11.1 and 28.7 +/- 14.6 pg/1O6 cells, respectively. When cells have been incubated for 2 to 72 hours with a P.E. inhibitor (Z-Gly-Pro-CHN2) at 5 x 10(-7) M, both cell PGP I and PE activities are inhibited. No change in the cellular content of immunoreactive TRH, His-Pro-Diketopiperazine and acid TRH have been observed in treated cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Published
1991

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