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Protein Disulfide Bond Formation in the Cytoplasm during Oxidative Stress

2004· article· en· 460 citations· W2044136163 on OpenAlex· 10.1074/jbc.m312267200

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Canadian funderA Canadian agency funded it. The work may carry no Canadian affiliation at all.

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Opus teacher head0.013
GPT teacher head0.251
Teacher spread
0.239 · how far apart the two teachers sit on this one work
Validation status
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Abstract

The majority of disulfide-linked cytosolic proteins are thought to be enzymes that transiently form disulfide bonds while catalyzing oxidation-reduction (redox) processes. Recent evidence indicates that reactive oxygen species can act as signaling molecules by promoting the formation of disulfide bonds within or between select redox-sensitive proteins. However, few studies have attempted to examine global changes in disulfide bond formation following reactive oxygen species exposure. Here we isolate and identify disulfide-bonded proteins (DSBP) in a mammalian neuronal cell line (HT22) exposed to various oxidative insults by sequential nonreducing/reducing two-dimensional SDS-PAGE combined with mass spectrometry. By using this strategy, several known cytosolic DSBP, such as peroxiredoxins, thioredoxin reductase, nucleoside-diphosphate kinase, and ribonucleotide-diphosphate reductase, were identified. Unexpectedly, a large number of previously unknown DSBP were also found, including those involved in molecular chaperoning, translation, glycolysis, cytoskeletal structure, cell growth, and signal transduction. Treatment of cells with a wide range of hydrogen peroxide concentrations either promoted or inhibited disulfide bonding of select DSBP in a concentration-dependent manner. Decreasing the ratio of reduced to oxidized glutathione also promoted select disulfide bond formation within proteins from cytoplasmic extracts. In addition, an epitope-tagged version of the molecular chaperone HSP70 forms mixed disulfides with both beta4-spectrin and adenomatous polyposis coli protein in the cytosol. Our findings indicate that disulfide bond formation within families of cytoplasmic proteins is dependent on the nature of the oxidative insult and may provide a common mechanism used to control multiple physiological processes.

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The record

Venue
Journal of Biological Chemistry
Topic
Redox biology and oxidative stress
Field
Biochemistry, Genetics and Molecular Biology
Canadian institutions
Funders
Canadian Institutes of Health ResearchNational Institutes of HealthJohn Douglas French Alzheimer's Foundation
Keywords
ChemistryProtein disulfide-isomeraseBiochemistryCytosolRibonucleotide reductaseOxidative stressOxidative phosphorylationGlutathione disulfideThioredoxinThioredoxin reductaseGlutathioneCytoplasmReactive oxygen speciesProtein subunitEnzyme
Has abstract in OpenAlex
yes