title: Proteomics analysis of cellular response to oxidative stress. Evidence for in vivo overoxidation of peroxiredoxins at their active site.
abstract: The proteomics analysis reported here shows that a major cellular response to oxidative stress is the modification of several peroxiredoxins. An acidic form of the peroxiredoxins appeared to be systematically increased under oxidative stress conditions. Peroxiredoxins are enzymes catalyzing the destruction of peroxides. In doing so, a reactive cysteine in the peroxiredoxin active site is weakly oxidized (disulfide or sulfenic acid) by the destroyed peroxides. Cellular thiols (e.g. thioredoxin) are used to regenerate the peroxiredoxins to their active state. Tandem mass spectrometry was carried out to characterize the modified form of the protein produced in vivo by oxidative stress. The cysteine present in the active site was shown to be oxidized into cysteic acid, leading to an inactivated form of peroxiredoxin. This strongly suggested that peroxiredoxins behave as a dam upon oxidative stress, being both important peroxide-destroying enzymes and peroxide targets. Results obtained in a primary culture of Leydig cells challenged with tumor necrosis factor alpha suggested that this oxidized/native balance of peroxiredoxin 2 may play an active role in resistance or susceptibility to tumor necrosis factor alpha-induced apoptosis.
authors.affiliation: CEA-Laboratoire de Bioénergétique Cellulaire et Pathologique, EA UJF 2943, DRDC/BECP, CEA-Grenoble, 17 rue des martyrs, F-38054 Grenoble Cedex 9, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Rabilloud
authors.forename: Manfred
authors.initials: M
authors.lastname: Heller
authors.forename: Francoise
authors.initials: F
authors.lastname: Gasnier
authors.forename: Sylvie
authors.initials: S
authors.lastname: Luche
authors.forename: Catherine
authors.initials: C
authors.lastname: Rey
authors.forename: Ruedi
authors.initials: R
authors.lastname: Aebersold
authors.forename: Mohamed
authors.initials: M
authors.lastname: Benahmed
authors.forename: Pierre
authors.initials: P
authors.lastname: Louisot
authors.forename: Joel
authors.initials: J
authors.lastname: Lunardi
mesh_terms: D000818:Animals; D017209:Apoptosis; D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry; D006801:Humans; D019169:Jurkat Cells; D013058:Mass Spectrometry; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D010544:Peroxidases / Q000737:chemistry* / Q000378:metabolism; D061105:Peroxiredoxin III; D054464:Peroxiredoxins; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D010455:Peptides; D014409:Tumor Necrosis Factor-alpha; D010544:Peroxidases; C516115:PRDX3 protein, human; D061105:Peroxiredoxin III; D054464:Peroxiredoxins; D003545:Cysteine; D010100:Oxygen