Submitted by Penny Peck on Tue, 09/12/2025 - 17:16
Hydrogen sulfide (H₂S) is increasingly recognised as an important signalling molecule in biological systems. It can reversibly modify low molecular weight (LMWSH) and protein (PrSH) thiols to form persulfides (RSS⁻) and polysulfides (RS(S)ₙS⁻), contributing to antioxidant defence and the regulation of protein function. However, progress in understanding the biological relevance of these modifications has been limited by the lack of methods to quantify them reliably in complex biological samples.
In a study led by Dr Jan Miljkovic and Professor Mike Murphy at the MRC Mitochondrial Biology Unit, in collaboration with colleagues from Professor Richard Hartley’s laboratory in Glasgow and partners in London and Cambridge, the team has developed a highly sensitive LC–MS/MS workflow that captures the sulfur atom of H₂S and the terminal sulfur atoms of RSS⁻ and RS(S)ₙS⁻ as diagnostic products in complex biological samples.
In parallel, they have established an approach to capture internal sulfur atoms in RS(S)ₙS⁻, enabling simultaneous quantification of H₂S, RSS⁻ and RS(S)ₙS⁻. Both LMWS(S)ₙS⁻ and PrS(S)ₙS⁻ are measured in the same sample. Glutathione (GSH) is the most abundant LMWSH so they then developed an orthogonal approach to quantify GSS-, enabling corroboration of LMWSS- measurements by sulfur atom trapping.
This work is published in Nature Communications, where the team demonstrates in systems from proteins to ex vivo tissues how these approaches enable exploration of persulfidation in biological systems.
Publication reference:
Miljkovic, J.L., Burger, N., Yu, C.S. et al. (2025) Simultaneous and sensitive quantification of protein and low molecular weight persulfides, polysulfides and H2S in biological samples. Nat Commun. https://doi.org/10.1038/s41467-025-66795-5