Quantitative Persulfide Site Identification (qPerS-SID) Reveals Protein Targets of H2S Releasing Donors in Mammalian Cells

• Published in: Scientific reports Vol. 6; no. 1; p. 29808
• Main Authors: Longen, Sebastian, Richter, Florian, Köhler, Yvette, Wittig, Ilka, Beck, Karl-Friedrich, Pfeilschifter, Josef
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 14.07.2016; Nature Publishing Group
• Subjects: 13; 13/106; 631/45/2783; 631/45/475; 631/80/458; 631/80/86; 82; Amino Acid Sequence; Amino acids; Biotin; Chromatography, Liquid - methods; Cysteine - analogs & derivatives; Cysteine - chemistry; Cysteine - metabolism; Disulfides - chemistry; Disulfides - metabolism; HEK293 Cells; Humanities and Social Sciences; Humans; Hydrogen sulfide; Hydrogen Sulfide - chemistry; Hydrogen Sulfide - metabolism; Inactivation; Kinases; Labeling; Mammals; Mass spectrometry; multidisciplinary; Peptides; Peptides - chemistry; Peptides - metabolism; Phosphatase; Proteins; Proteome - chemistry; Proteome - metabolism; Proteomics; Proteomics - methods; Reagents; Science; Scientific imaging; Sulfides - chemistry; Sulfides - metabolism; Tandem Mass Spectrometry - methods
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep29808

H 2 S 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 H 2 S 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 H 2 S signalling and enlightens the biology of H 2 S and its releasing agents.