Dataset Information

Redox proteome analysis identifies reversible oxidized proteins in retinoic acid differentiated SH-SY5Y cells

ABSTRACT: Retinoic acid stimulated SH-SY5Y cells are a common model system to study the initial phases of neuronal differentiation. Although these processes are associated with redox-sensitive processes, no comprehensive analysis of oxidative modified cysteines, as one of the main targets for reactive oxygen species, was available. Here, we used quantitative and differential proteomics and redox proteomics, to fill this gap. Our results indicate, that alterations in the redox state very early in the differentiation process, affect the expression of marker proteins and the extension of neurites. The spatiotemporal analysis of reactive oxygen species, suggests a NOX-dependent peak in cytoplasmic O2.-/H2O2 level 2h after retinoic acid stimulation. At the same timepoint 241 out of 275 proteins with an altered cysteine redox state appeared to be reversibly oxidised in the retinoic acid treated samples. Our analyses suggest that redox alterations affect proteins involved in the retinoic acid homeostasis and cytoskeletal dynamics.

INSTRUMENT(S): LTQ Orbitrap Elite, Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Neuroblast

SUBMITTER: Gereon Poschmann

LAB HEAD: Gereon Poschmann

PROVIDER: PXD014381 | Pride | 2020-03-30

REPOSITORIES: Pride

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Publications

The Proteomic Landscape of Cysteine Oxidation That Underpins Retinoic Acid-Induced Neuronal Differentiation.

Brenig Katrin K Grube Leonie L Schwarzländer Markus M Köhrer Karl K Stühler Kai K Poschmann Gereon G

Journal of proteome research 20200403 5

The initial phases of neuronal differentiation are key to neuronal function. A particularly informative model to study these initial phases are retinoic acid-stimulated SH-SY5Y cells. Although these progressions are associated with redox-sensitive processes, it is largely undefined how the cellular proteome underpins redox dynamics and the management of reactive oxygen species. Here, we map the global cysteine-based redox landscape of SH-SY5Y cells using quantitative redox proteomics. We find ev ...[more]

PMID: 32202429

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Project description:Oxidative stress due to endogenous hydrogen peroxide production by Lactobacillus species is a well-known issue in the food industry. In this study, the transcriptional response to oxygen of Lactobacillus johnsonii, one of the H2O2-producing strains used in the food industry, was analyzed. It was found that aerobic growth conditions led to a more than two-fold downregulation of 45 genes as compared to anaerobic growth, whereas 6 genes were more than twofold upregulated. Among the upregulated genes were two genes that displayed significant homology to NADH-dependent oxidoreductase (NOX). The postulated transcriptional regulation of the nox promoter by oxygen was studied using a GUS-reporter construct, confirming a 2.1-fold upregulated GUS-expression upon aerobic growth. Exposure to sublethal levels of hydrogen peroxide did not result in significant regulation of the nox promoter. In a previous study of hydrogen peroxide production by L. johnsonii, a NADH flavin reductase (NFR) was identified to be involved in hydrogen peroxide production. An NFR-deficient derivative was strongly impaired in H2O2 production, but regained a partial H2O2 producing capacity upon prolonged oxygen exposure. The nox-promoter appeared to be 3.6-fold upregulated under aerobic conditions in the NFR-deficient background, which may imply a role of this gene in the regained H2O2 production. Indeed, deletion of the nox-gene in the NFR-deletion background, resulted in a strain that no longer produced H2O2, also during prolonged exposure to oxygen. The double-mutant (nfr, nox) displayed strongly impaired aerobic growth and oxygenation induced rapid growth stagnation that is not caused by H2O2. We conclude that H2O2 production in L. johnsonii is primarily dependent on NFR but can also involve an oxygen-inducible NADH oxidase under aerobic conditions. Moreover, our results imply that H2O2 production plays a prominent role in oxygen tolerance of L. johnsonii. loop design of the samples including two shortcuts

Dataset Information

Redox proteome analysis identifies reversible oxidized proteins in retinoic acid differentiated SH-SY5Y cells

Publications

The Proteomic Landscape of Cysteine Oxidation That Underpins Retinoic Acid-Induced Neuronal Differentiation.

Similar Datasets

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