{"doi":"10.1101/2023.03.10.532002","title":"Cryo-EM structures of human arachidonate 12S-Lipoxygenase (12-LOX) bound to endogenous and exogenous inhibitors","abstract":"Abstract Human 12-lipoxygenase (12-LOX) is an enzyme involved in platelet activation and is a promising target for antiplatelet therapies. Despite the clinical importance of 12-LOX, the exact mechanisms of how it affects platelet activation are unclear, and the lack of structural information has limited drug discovery efforts. In this study, we used single-particle cryoelectron microscopy to determine the high-resolution structures (1.7 Å - 2.8 Å) of human 12-LOX for the first time. Our results showed that 12-LOX can exist in multiple oligomeric states, from monomer to hexamer, which may impact its catalytic activity and membrane association. We also identified different conformations within a 12-LOX dimer, likely representing different time points in its catalytic cycle. Furthermore, we were able to identify small molecules bound to the 12-LOX structures. The active site of the 12-LOX tetramer is occupied by an endogenous 12-LOX inhibitor, a long-chain acyl-Coenzyme A. Additionally, we found that the 12-LOX hexamer can simultaneously bind to arachidonic acid and ML355, a selective 12-LOX inhibitor that has passed a phase I clinical trial for treating heparin-induced thrombocytopenia and has received fast-track designation by the FDA. Overall, our findings provide novel insights into the assembly of 12-LOX oligomers, its catalytic mechanism, and small molecule binding, paving the way for further drug development targeting the 12-LOX enzyme. Key Points The first full-length structures of human arachidonate 12S-Lipoxygenase (12-LOX) Reveals mechanisms of oligomeric and conformational states Uncovers natural inhibitor of 12S-Lipoxygenase (12-lox) Reveals a binding site of inhibitor ML355","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":395974,"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":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9434,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1039108,"name":"Katrina A. Black","orcid":"0000-0002-4094-6170","position":1,"is_corresponding":false},{"id":966682,"name":"Michelle Tran","orcid":"0000-0002-7578-9347","position":2,"is_corresponding":false},{"id":347611,"name":"Hariprasad Venugopal","orcid":"0000-0001-5230-2973","position":3,"is_corresponding":false},{"id":329819,"name":"Theodore R. Holman","orcid":"0000-0001-8072-2959","position":4,"is_corresponding":false},{"id":277616,"name":"Michael Holinstat","orcid":"0000-0001-5100-1933","position":5,"is_corresponding":false},{"id":638378,"name":"David M. Thal","orcid":"0000-0002-0325-2524","position":6,"is_corresponding":false},{"id":307917,"name":"Alisa Glukhova","orcid":"0000-0003-4146-965X","position":7,"is_corresponding":false},{"id":641075,"name":"Jesse I. Mobbs","orcid":"0000-0001-6979-1427","position":0,"is_corresponding":true}],"reference_count":51,"raw_metadata":null,"created_at":"2026-07-19T01:19:27.044553Z","pmid":null,"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":[]}