{"doi":"10.3390/biom15030402","title":"The Eye Lens Protein, γS Crystallin, Undergoes Glutathionylation-Induced Disulfide Bonding Between Cysteines 22 and 26","abstract":"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 γS-crystallin to glutathionylation-induced oxidation and disulfide bond formation. Recombinantly expressed wild-type human γS-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, γS-crystallin undergoes stepwise oxidation reactions and disulfide shuffling, which may contribute towards its aggregation and cataract formation.","journal":"Biomolecules","year":2025,"id":519202,"datarank":0.29188652235829704,"base_score":1.9459101490553132,"endowment":1.9459101490553132,"self_citation_contribution":0.29188652235829704,"citation_network_contribution":0.0,"self_endowment_contribution":0.29188652235829704,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":6,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9577,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":569756,"name":"David C. Thorn","orcid":"0000-0002-7332-2292","position":1,"is_corresponding":false},{"id":1126023,"name":"Martin E. Tovar‐Ramírez","orcid":"0000-0002-3186-3808","position":2,"is_corresponding":false},{"id":247329,"name":"Eugene I. Shakhnovich","orcid":"0000-0002-4769-2265","position":3,"is_corresponding":false},{"id":141412,"name":"Larry L. David","orcid":null,"position":4,"is_corresponding":false},{"id":141387,"name":"Kirsten J. Lampi","orcid":"0000-0002-7906-6699","position":5,"is_corresponding":false},{"id":1281935,"name":"Kate Halverson-Kolkind","orcid":null,"position":0,"is_corresponding":true}],"reference_count":33,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:49:18.751199Z","pmid":"40149938","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}