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The legacy of a persistent role for H2S in life\u2019s processes recently emerged through its discovery in modern biochemistry as an endogenous cellular signalling modulator involved in numerous biological processes. One major mechanism through which H2S signals is protein cysteine persulfidation, an oxidative post-translational modification. In recent years, chemoproteomic technologies have been developed to allow the global scanning of protein persulfidation targets in mammalian cells and tissues, providing a powerful tool to elucidate the broader impact of altered H2S in organismal physiological health and human disease states. While hundreds of proteins were confirmed to be persulfidated by global persulfidome methodologies, the targeting of specific proteins of interest and the investigation of further mechanistic studies are still underdeveloped due to a lack of stringent specificity of the methods and the inherent instability of persulfides. This review provides an overview of the processes of endogenous H2S production, oxidation, and signalling and highlights the application and limitations of current persulfidation labelling approaches for investigation of this important evolutionarily conserved biological switch for protein function.<\/jats:p>","DOI":"10.1042\/ebc20230095","type":"journal-article","created":{"date-parts":[[2024,9,18]],"date-time":"2024-09-18T07:39:34Z","timestamp":1726645174000},"page":"467-478","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Quantification of persulfidation on specific proteins: are we nearly there yet?"],"prefix":"10.1042","volume":"68","author":[{"given":"Hongling","family":"Liu","sequence":"additional","affiliation":[{"name":"1Molecular Metabolism Group, University\/BHF Centre for Cardiovascular Sciences, Queens Medical Research Institute, University of Edinburgh, U.K."},{"name":"2Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen 2200, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Florentina","family":"Negoita","sequence":"additional","affiliation":[{"name":"2Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen 2200, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew","family":"Brook","sequence":"additional","affiliation":[{"name":"1Molecular Metabolism Group, University\/BHF Centre for Cardiovascular Sciences, Queens Medical Research Institute, University of Edinburgh, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8839-5980","authenticated-orcid":false,"given":"Kei","family":"Sakamoto","sequence":"additional","affiliation":[{"name":"2Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen 2200, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8218-8462","authenticated-orcid":false,"given":"Nicholas\u00a0M.","family":"Morton","sequence":"additional","affiliation":[{"name":"1Molecular Metabolism Group, University\/BHF Centre for Cardiovascular Sciences, Queens Medical Research Institute, University of Edinburgh, U.K."},{"name":"3Centre for Systems Health and Integrated Metabolic Research, Department of Biosciences, School of Science and Technology, Nottingham Trent University, NG11 8NS, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2024,12,4]]},"reference":[{"key":"2024120413260495300_B1","doi-asserted-by":"publisher","first-page":"5526","DOI":"10.1073\/pnas.1019191108","article-title":"Primordial synthesis of amines and amino acids in a 1958 Miller H2S-rich spark discharge experiment","volume":"108","author":"Parker","year":"2011","journal-title":"Proc. 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