Quantitative persulfide site identification (qPerS-SID) reveals protein targets of H2S releasing donors in mammalian cells

  • H2S is an important signalling molecule involved in diverse biological processes. It mediates the formation of cysteine persulfides (R-S-SH), which affect the activity of target proteins. Like thiols, persulfides show reactivity towards electrophiles and behave similarly to other cysteine modifications in a biotin switch assay. In this manuscript, we report on qPerS-SID a mass spectrometry-based method allowing the isolation of persulfide containing peptides in the mammalian proteome. With this method, we demonstrated that H2S donors differ in their efficacy to induce persulfides in HEK293 cells. Furthermore, data analysis revealed that persulfide formation affects all subcellular compartments and various cellular processes. Negatively charged amino acids appeared more frequently adjacent to cysteines forming persulfides. We confirmed our proteomic data using pyruvate kinase M2 as a model protein and showed that several cysteine residues are prone to persulfide formation finally leading to its inactivation. Taken together, the site-specific identification of persulfides on a proteome scale can help to identify target proteins involved in H2S signalling and enlightens the biology of H2S and its releasing agents.
Author:Sebastian Longen, Florian Richter, Yvette Köhler, Ilka WittigORCiD, Karl-Friedrich Beck, Josef Martin PfeilschifterGND
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/27411966
Parent Title (English):Scientific reports
Publisher:Macmillan Publishers Limited, part of Springer Nature
Place of publication:[London]
Document Type:Article
Year of Completion:2016
Date of first Publication:2016/07/14
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2018/06/07
Tag:Cell signalling; Chemical modification; Post-translational modifications; Proteomics
Issue:Art. 29808
Page Number:12
First Page:1
Last Page:12
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Institutes:Medizin / Medizin
Wissenschaftliche Zentren und koordinierte Programme / Sonderforschungsbereiche / Forschungskollegs
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Licence (German):License LogoCreative Commons - Namensnennung 4.0