Insight into protein S-nitrosylation in Chlamydomonas reinhardtii.

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The importance of nitrosylation in photosynthetic eukaryotes has been less studied. The aim of this study was to expand our knowledge on protein nitrosylation by performing a large-scale proteomic analysis of proteins undergoing nitrosylation in vivo in Chlamydomonas reinhardtii cells under nitrosative stress.\nRESULTS: Using two complementary proteomic approaches, 492 nitrosylated proteins were identified. They participate in a wide range of biological processes and pathways, including photosynthesis, carbohydrate metabolism, amino acid metabolism, translation, protein folding or degradation, cell motility, and stress. Several proteins were confirmed in vitro by western blot, site-directed mutagenesis and activity measurements. Moreover, 392 sites of nitrosylation were also identified. These results strongly suggest that S-nitrosylation could constitute a major mechanism of regulation in C. reinhardtii under nitrosative stress conditions.\nINNOVATION: This study constitutes the largest proteomic analysis of protein nitrosylation reported to date.\nCONCLUSION: The identification of 381 previously unrecognized targets of nitrosylation further extends our knowledge on the importance of this PTM in photosynthetic eukaryotes. The data have been deposited to the ProteomeXchange repository with identifier PXD000569.", "claims": [{"id": "claim-target-result", "statement": "The paper used two complementary proteomic approaches, and the source data were deposited in the ProteomeXchange repository under identifier PXD000569.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:24328795", "locator": "sections[0] \"Abstract\" paragraph 1", "quote": "Using two complementary proteomic approaches, 492 nitrosylated proteins were identified."}, {"asset": "source:pmid:24328795", "locator": "sections[0] \"Abstract\" paragraph 1", "quote": "The data have been deposited to the ProteomeXchange repository with identifier PXD000569."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": "The paper used two complementary proteomic approaches, and the source data were deposited in the ProteomeXchange repository under identifier PXD000569.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:24328795"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "blocked", "blocked_on": "data:accession_absent", "description": "The GEO series accession resolves and its sample accessions are listed. 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