Essential Role of Methionine Residues in Calmodulin Binding to Bordetella pertussis Adenylate Cyclase, as Probed by Selective Oxidation and Repair by the Peptide Methionine Sulfoxide Reductases
| Autor: | Daniel Ladant, Nathalie Dautin, Stéphanie Vougier, Jean Yves Mary, Joëlle Vinh, Bertrand Friguet |
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| Přispěvatelé: | Biologie et Biochimie Cellulaire du Vieillissement ((EA_3106)), Université Paris Diderot - Paris 7 (UPD7), Biochimie des Interactions Macromoléculaires, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Neurobiologie et diversité cellulaire (NDC), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2004 |
| Předmět: |
Bordetella pertussis
Neutrophils [SDV]Life Sciences [q-bio] MESH: Adenylyl Cyclases Plasma protein binding MESH: Neutrophils Biochemistry Mass Spectrometry MESH: Dose-Response Relationship Drug MESH: Recombinant Proteins MESH: Bordetella pertussis chemistry.chemical_compound MESH: Protein Structure Tertiary Methionine tert-Butylhydroperoxide Cyclic AMP MESH: Animals MESH: Phagocytosis MESH: Cyclic AMP 0303 health sciences CD11b Antigen biology MESH: Kinetics MESH: Escherichia coli MESH: Peptides Microfilament Proteins 030302 biochemistry & molecular biology MESH: Enzyme-Linked Immunosorbent Assay MESH: Transcription Factors MESH: Calmodulin Recombinant Proteins MESH: Surface Plasmon Resonance MESH: Methionine Sulfoxide Reductases Methionine sulfoxide reductase Electrophoresis Polyacrylamide Gel Oxidoreductases MESH: Oxygen MESH: Spectrometry Fluorescence Adenylyl Cyclases Plasmids Protein Binding MSRA Spectrometry Mass Electrospray Ionization MESH: Ions Calmodulin MESH: Rats Adenylate kinase Enzyme-Linked Immunosorbent Assay MESH: CD11b Antigen Cyclase MESH: Spectrometry Mass Electrospray Ionization 03 medical and health sciences Phagocytosis MESH: Plasmids Escherichia coli Animals Humans MESH: Protein Binding [CHIM]Chemical Sciences Biotinylation MESH: Biotinylation MESH: Oxidoreductases Molecular Biology 030304 developmental biology Ions MESH: Mass Spectrometry MESH: Humans Dose-Response Relationship Drug Macrophages MESH: Macrophages Cell Biology Surface Plasmon Resonance biology.organism_classification Protein Structure Tertiary Rats Oxygen Kinetics Spectrometry Fluorescence chemistry MESH: Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization CD18 Antigens Methionine Sulfoxide Reductases Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization MESH: Methionine MESH: tert-Butylhydroperoxide biology.protein Peptides MESH: CD18 Antigens Transcription Factors MESH: Electrophoresis Polyacrylamide Gel |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2004, 279 (29), pp.30210-30218. ⟨10.1074/jbc.M400604200⟩ Journal of Biological Chemistry, 2004, 279 (29), pp.30210-30218. ⟨10.1074/jbc.M400604200⟩ |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1074/jbc.M400604200⟩ |
| Popis: | International audience; Bordetella pertussis, the causative agent of whooping cough, secretes among other virulence factors an adenylate cyclase (AC) toxin that is able to enter into eukaryotic cells where it is activated upon binding to endogenous calmodulin (CaM) and synthesizes supraphysiological cAMP levels. In vivo, the AC toxin, through its specific interaction with the CD11b/CD18 integrin, primarily targets phagocytic cells such as neutrophils and macrophages. Because neutrophil priming and activation result in the production of reactive oxygen species that may cause intracellular oxidation, we have examined the biological consequences of the oxidation of CaM methionines upon its interaction with AC. We show here that the interaction of CaM with AC is dependent on the reduced state of methionines, because oxidation of all methionine residues of CaM dramatically decreases its affinity for AC. Peptide methionine sulfoxide reductases A (MsrA) and B (MsrB) were able to partially reduce the oxidized CaM, and these partially "repaired" forms could interact with AC nearly as efficiently as the native protein. We further showed that the CaM.AC complex is resistant to oxidation with tert-butylhydroperoxide, and we identified methionine residues 109, 124, and 145 as critical for binding to AC. The resistance of the AC.CaM complex to oxidation and the ability of AC to be efficiently activated by partially oxidized CaM molecules should allow the toxin to exert its cytotoxic effects on activated neutrophils and contribute to the host colonization. |
| Databáze: | OpenAIRE |
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