{"doi":"10.1002/pro.70344","title":"Atomistic modulation of <scp>MIF</scp> ‐2 structure, catalysis, and biological signaling via cysteine residues and a small molecule, Ebselen","abstract":"The macrophage migration inhibitory factor (MIF) family of cytokines comprised of the MIF and D-dopachrome tautomerase (or MIF-2) paralogs share identical tertiary and quaternary structures that contribute to their overlapping enzymatic and signaling activities. Recent investigations of MIF and MIF-2 have shown them to possess N-to-C-terminal allosteric crosstalk, but despite the similarity of this \"allosteric pathway,\" its regulation of MIF and MIF-2 is not identical. Thus, structure alone does not preserve the precise allosteric mechanism, and additional residues that modulate MIF and MIF-2 function must be characterized. Cysteines have been identified as allosteric switches for the same biochemical functions of MIF, and small molecules targeting its N-terminal enzymatic site have affected the structure of three proximal cysteines. Ebselen is a compound that forms covalent selenylsulfide bonds with MIF cysteines and is hypothesized to destabilize and dissociate the MIF trimer into monomers. Ebselen-bound MIF also displays little-to-no catalysis or biological signaling. However, it is unclear whether Ebselen similarly affects the MIF-2 paralog, despite MIF-2 containing two related cysteines (MIF contains three). We used mutagenesis, nuclear magnetic resonance, molecular dynamics simulations, in vitro and in vivo biochemistry to investigate the mechanism of Ebselen as a modulator of MIF-2 via its cysteines. Our findings suggest that Ebselen partially disrupts the MIF-2 homotrimer, though the overall population of such a structure is <35%, even on the timescale of many hours. Ebselen does attenuate the biological functions of MIF-2, and solution structural biology captures the conformational transitions preceding the destabilized MIF-2 trimer.","journal":"Protein Science","year":2025,"id":547739,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9545,"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":1440964,"name":"Sirena M. D'Orazio","orcid":null,"position":1,"is_corresponding":false},{"id":424665,"name":"Pragnya Das","orcid":"0000-0003-4818-2387","position":2,"is_corresponding":false},{"id":806923,"name":"Divya Rajendran","orcid":"0000-0002-5860-9586","position":3,"is_corresponding":false},{"id":1440965,"name":"Xander Takada","orcid":null,"position":4,"is_corresponding":false},{"id":913882,"name":"Yuanjun Shi","orcid":null,"position":5,"is_corresponding":false},{"id":1440966,"name":"Iz Varghese","orcid":null,"position":6,"is_corresponding":false},{"id":951864,"name":"Yannie Lam","orcid":"0009-0003-9472-2679","position":7,"is_corresponding":false},{"id":476516,"name":"Nicholas A. DaSilva","orcid":"0000-0003-1925-0790","position":8,"is_corresponding":false},{"id":539578,"name":"Jimin Wang","orcid":"0000-0002-4504-8038","position":9,"is_corresponding":false},{"id":340256,"name":"Víctor S. Batista","orcid":"0000-0002-3262-1237","position":10,"is_corresponding":false},{"id":339237,"name":"Vineet Bhandari","orcid":"0000-0002-4900-6636","position":11,"is_corresponding":false},{"id":340257,"name":"George P. Lisi","orcid":"0000-0001-8878-5655","position":12,"is_corresponding":false},{"id":739606,"name":"Vinnie Widjaja","orcid":"0000-0002-3190-5633","position":0,"is_corresponding":true}],"reference_count":62,"raw_metadata":null,"created_at":"2026-07-19T02:53:53.962932Z","pmid":"41099614","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":[]}