{"doi":"10.1016/j.mcpro.2025.101315","title":"Expanding the Crosslinking Mass Spectrometry Toolbox With Vinyl Sulfone Crosslinkers","abstract":"Crosslinking mass spectrometry (MS) is a powerful approach for probing protein structures. However, most widely used crosslinkers rely on N-hydroxysuccinimide (NHS) esters, restricting reactivity primarily to lysine residues and protein N-termini, and rendering them incompatible with many amine-containing buffers (e.g., Tris) and key biochemical cofactors (e.g., ATP). To address these limitations, we introduce two novel vinyl-sulfone-based crosslinkers. Alkyne-BVSC is an enrichable, homobifunctional crosslinker featuring an acid-cleavable alkyne handle for downstream peptide enrichment. VSD is a heterobifunctional crosslinker combining a vinyl sulfone with a diazirine moiety for UV-activated photo-crosslinking. Both reagents are synthetically accessible from inexpensive precursors and retain reactivity in amine-rich biochemical environments. We show that vinyl sulfones react with cysteine, histidine, and lysine residues, thereby expanding crosslinkable residues beyond those accessible to NHS-esters. Moreover, we develop a stub-based post-search filtering strategy that leverages the MS-cleavable nature of vinyl sulfone linkages to improve crosslink identification sensitivity. Together, these advances establish vinyl-sulfone-based crosslinkers as versatile and complementary tools for structural proteomics.","journal":"Molecular & Cellular Proteomics","year":2025,"id":578738,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9602,"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":772756,"name":"Haitao Wu","orcid":"0000-0002-7617-2832","position":1,"is_corresponding":false},{"id":1455843,"name":"Katerina Atallah-Yunes","orcid":"0009-0001-2530-1729","position":2,"is_corresponding":false},{"id":1031227,"name":"Kitaik Lee","orcid":"0000-0002-7761-5586","position":3,"is_corresponding":false},{"id":1489003,"name":"Chris Sibley","orcid":null,"position":4,"is_corresponding":false},{"id":959369,"name":"Jesse A. Spivey","orcid":"0000-0001-7001-422X","position":5,"is_corresponding":false},{"id":663568,"name":"Rolf E. Swenson","orcid":"0000-0001-7804-7202","position":6,"is_corresponding":false},{"id":249547,"name":"John S. Schneekloth","orcid":"0000-0001-7459-783X","position":7,"is_corresponding":false},{"id":655109,"name":"Francis J. O’Reilly","orcid":"0000-0001-9258-0150","position":8,"is_corresponding":false},{"id":713353,"name":"Anthony M. Ciancone","orcid":"0000-0002-6240-1542","position":0,"is_corresponding":true}],"reference_count":45,"raw_metadata":null,"created_at":"2026-07-19T02:58:24.957414Z","pmid":"41130296","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":[]}