Proteomics Analysis of Cellular Response to Oxidative Stress: EVIDENCE FOR IN VIVO OVEROXIDATION OF PEROXIREDOXINS AT THEIR ACTIVE SITE

Thierry Rabilloud,Manfred Heller,Françoise Gasnier,Sylvie Luche,Catherine Rey,Ruedi Aebersold,Mohamed Benahmed,Pierre Louisot,Joël Lunardi

doi:10.1074/jbc.m106585200

Thierry Rabilloud, Manfred Heller + Show 7 more

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https://doi.org/10.1074/jbc.m106585200

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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.

Highlights

The proteomics analysis reported here shows that a major cellular response to oxidative stress is the modification of several peroxiredoxins
Results obtained in a primary culture of Leydig cells challenged with tumor necrosis factor ␣ suggested that this oxidized/native balance of peroxiredoxin 2 may play an active role in resistance or susceptibility to tumor necrosis factor ␣-induced apoptosis
Acidic Peroxiredoxin Spots Appear upon Oxidative Stress—In a search for protein modification occurring after oxidative stress, we used a proteomic approach on Jurkat cells stressed mildly with glucose oxidase or strongly with butyl hydroperoxide

Summary

Introduction

The proteomics analysis reported here shows that a major cellular response to oxidative stress is the modification of several peroxiredoxins. 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 peroxidedestroying enzymes and peroxide targets. Organisms living under aerobic conditions need to protect themselves against the damage caused by reactive oxygen species (O2., H2O2, and OH1⁄7), arising from either the incomplete reduction of oxygen during cellular respiration or exposure to external agents such as light, ionizing radiation, or some redox drugs [1, 2] These reactive oxygen species can damage various components of living cells such as unsaturated lipids (giving rise to deleterious organic peroxides), proteins, or nucleic acids. To complete the enzymatic catalytic cycle, the peroxiredoxins are reduced back to their active thiol form, for example by the thioredoxin-thioredoxin reductase system for 2-Cys peroxiredoxins [5, 6]

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