Simultaneous and sensitive quantification of protein and low molecular weight persulfides, polysulfides and H<sub>2</sub>S in biological samples.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 41345365.
- Also identified by DOI 10.1038/s41467-025-66795-5 and PMC identifier 12769671.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
H<sub>2</sub>S reversibly modifies low molecular weight (L<sub>MW</sub>SH) and protein (PrSH) thiols to form persulfides (RSS<sup>-</sup>) and polysulfides (RS(S)<sub>n</sub>S<sup>-</sup>) for antioxidant defence and regulation of activity. However, our understanding of the biological significance of these processes is hampered by our inability to quantify these modifications. We develop a sensitive LC-MS/MS procedure that traps the sulfur atom of H<sub>2</sub>S, and the terminal sulfur atom of RSS<sup>-</sup> and RS(S)<sub>n</sub>S<sup>-</sup> as diagnostic products in biological samples. In parallel, we also trap internal S atoms of RS(S)<sub>n</sub>S<sup>-</sup>, enabling quantification of H<sub>2</sub>S, RSS<sup>-</sup> and RS(S)<sub>n</sub>S<sup>-</sup>. L<sub>MW</sub>S(S)<sub>n</sub>S<sup>-</sup> and PrS(S)<sub>n</sub>S<sup>-</sup> are determined simultaneously in the same sample. Glutathione (GSH) is the most abundant L<sub>MW</sub>SH so we develop an orthogonal approach to quantify GSS<sup>-</sup>, enabling corroboration of L<sub>MW</sub>SS<sup>-</sup> measurements by sulfur atom trapping. We demonstrate in systems from proteins to ex vivo tissues how these approaches enable exploration of persulfidation in biological systems.
Medical subject headings
- Hydrogen Sulfide
- Sulfides
- Proteins