{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T11:17:45Z","timestamp":1785237465918,"version":"3.55.0"},"reference-count":35,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,3,11]],"date-time":"2025-03-11T00:00:00Z","timestamp":1741651200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH-National Eye Institute","award":["R01EY027768"],"award-info":[{"award-number":["R01EY027768"]}]},{"name":"NIH-National Eye Institute","award":["R01EY030444"],"award-info":[{"award-number":["R01EY030444"]}]},{"name":"NIH-National Eye Institute","award":["R01EY027012"],"award-info":[{"award-number":["R01EY027012"]}]},{"name":"NIH-National Eye Institute","award":["P30EY010572"],"award-info":[{"award-number":["P30EY010572"]}]},{"name":"NIH-National Eye Institute","award":["S10OD012246"],"award-info":[{"award-number":["S10OD012246"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>The oxidation of cysteines in crystallins is a major age-related modification associated with cataract formation. The purpose of this research was to determine the susceptibility of \u03b3S-crystallin to glutathionylation-induced oxidation and disulfide bond formation. Recombinantly expressed wild-type human \u03b3S-crystallin and four cysteine-to-serine mutants were reduced and incubated for up to 2 days with oxidized glutathione. Following incubation and alkylation, the overall degree of glutathionylation and disulfide bond formation were determined by whole-mass measurement. Tryptic digests were also analyzed by LC-MS\/MS to identify specific sites of S-glutathionylation and disulfide linkages. We determined that C22, C24, and C26 undergo glutathione-mediated disulfide interchange with each other, with C24 being most susceptible to oxidation and mixed disulfide formation. Our data suggest C24 is S-glutathionylated sequentially with C22 and C26 participating in disulfide exchange reactions, yielding a major species with a single glutathionylation at C24 and a disulfide between C22 and C26. The results imply that as glutathione levels are depleted in aged lenses, \u03b3S-crystallin undergoes stepwise oxidation reactions and disulfide shuffling, which may contribute towards its aggregation and cataract formation.<\/jats:p>","DOI":"10.3390\/biom15030402","type":"journal-article","created":{"date-parts":[[2025,3,11]],"date-time":"2025-03-11T08:59:52Z","timestamp":1741683592000},"page":"402","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["The Eye Lens Protein, \u03b3S Crystallin, Undergoes Glutathionylation-Induced Disulfide Bonding Between Cysteines 22 and 26"],"prefix":"10.3390","volume":"15","author":[{"given":"Kate","family":"Halverson-Kolkind","sequence":"first","affiliation":[{"name":"Biomaterials and Biomedical Science, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7332-2292","authenticated-orcid":false,"given":"David","family":"Thorn","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3186-3808","authenticated-orcid":false,"given":"Martin","family":"Tovar-Ramirez","sequence":"additional","affiliation":[{"name":"Biomaterials and Biomedical Science, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eugene","family":"Shakhnovich","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Larry","family":"David","sequence":"additional","affiliation":[{"name":"Chemical Physiology & Biochemistry, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7906-6699","authenticated-orcid":false,"given":"Kirsten","family":"Lampi","sequence":"additional","affiliation":[{"name":"Biomaterials and Biomedical Science, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1001\/archopht.122.4.487","article-title":"Prevalence of cataract and pseudophakia\/aphakia among adults in the United States","volume":"122","author":"Congdon","year":"2004","journal-title":"Arch. 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