This report characterizes and quantifies endogenous hydrogen sulfide (H₂S) and small oxoacids of sulfur (SOS = HOSH, HOSOH) in a panel of cell lines including human cancer (A375 melanoma cells, HeLa cervical cells) and noncancer (HEK293 embryonic kidney cells), as well as E. coli DH5α and S. cerevisiae S288C. The methodology used is a translation of well-studied nucleophilic and electrophilic traps for cysteine and oxidized cysteines residues to target small molecular weight sulfur species; mass spectrometric analysis allows for species quantification. The observed intracellular concentrations of H₂S and SOS vary in different cell types, from nanomolar to femtomolar, typically with H₂S > HOSOH > HOSH. We propose the term sulfome, a subset of the metabolome, describing the nonproteinaceous metabolites of H₂S; the sulfomic index is as a measure of the S-oxide redox status, which gives a profile of endogenous sulfur at different oxidation states. An important observation is that H₂S and SOS were found to be continuously extruded into surrounding media against a concentration gradient, implying an active efflux process. Small molecule inhibition of several H₂S generating enzymes suggest that SOS are not derived solely from H₂S oxidation. Even after successful inhibition of H₂S production, cells maintain constant efflux and repopulate H₂S and SOS over time. This work proves that these small sulfur oxoacids are generated in cells of all types, and their efflux implies that they play a role in cell signaling and possibly other vascular physiology attributed to H₂S.

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Journal Article | Research Support, N.I.H., Extramural | Research Support, Non-U.S. Gov't

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Scrivner O, Kumar MR, Sorokolet K, Wong A, Kebaara B, Farmer PJ

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