{"doi":"10.3389/fphar.2022.891041","title":"Screening of Hydrocarbon-Stapled Peptides for Inhibition of Calcium-Triggered Exocytosis","abstract":"The so-called primary interface between the SNARE complex and synaptotagmin-1 (Syt1) is essential for Ca 2+ -triggered neurotransmitter release in neuronal synapses. The interacting residues of the primary interface are conserved across different species for synaptotagmins (Syt1, Syt2, Syt9), SNAP-25, and syntaxin-1A homologs involved in fast synchronous release. This Ca 2+ -independent interface forms prior to Ca 2+ -triggering and plays a role in synaptic vesicle priming. This primary interface is also conserved in the fusion machinery that is responsible for mucin granule membrane fusion. Ca 2+ -stimulated mucin secretion is mediated by the SNAREs syntaxin-3, SNAP-23, VAMP8, Syt2, and other proteins. Here, we designed and screened a series of hydrocarbon-stapled peptides consisting of SNAP-25 fragments that included some of the key residues involved in the primary interface as observed in high-resolution crystal structures. We selected a subset of four stapled peptides that were highly α-helical as assessed by circular dichroism and that inhibited both Ca 2+ -independent and Ca 2+ -triggered ensemble lipid-mixing with neuronal SNAREs and Syt1. In a single-vesicle content-mixing assay with reconstituted neuronal SNAREs and Syt1 or with reconstituted airway SNAREs and Syt2, the selected peptides also suppressed Ca 2+ -triggered fusion. Taken together, hydrocarbon-stapled peptides that interfere with the primary interface consequently inhibit Ca 2+ -triggered exocytosis. Our inhibitor screen suggests that these compounds may be useful to combat mucus hypersecretion, which is a major cause of airway obstruction in the pathophysiology of COPD, asthma, and cystic fibrosis.","journal":"Frontiers in Pharmacology","year":2022,"id":280783,"datarank":0.47761269701238607,"base_score":2.302585092994046,"endowment":2.302585092994046,"self_citation_contribution":0.3453877639491069,"citation_network_contribution":0.13222493306327912,"self_endowment_contribution":0.3453877639491069,"citer_contribution":0.13222493306327912,"corpus_percentile":null,"corpus_rank":null,"citation_count":9,"citer_count":5,"citers_with_citation_signal":4,"citers_with_endowment":4,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9494,"is_data_producer":true,"deposit_databanks":{"Dryad":["10.5061/dryad.x3ffbg7mp"]},"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":364805,"name":"Michael J. Tuvim","orcid":"0000-0001-8126-7716","position":1,"is_corresponding":false},{"id":749935,"name":"Jeremy Leitz","orcid":"0009-0009-9976-2497","position":2,"is_corresponding":false},{"id":749934,"name":"John Jacob Peters","orcid":"0000-0002-3192-7824","position":3,"is_corresponding":false},{"id":51694,"name":"Richard A. Pfuetzner","orcid":"0000-0002-1741-2330","position":4,"is_corresponding":false},{"id":299248,"name":"Luis Esquivies","orcid":"0000-0002-1750-6775","position":5,"is_corresponding":false},{"id":299246,"name":"Qiangjun Zhou","orcid":"0000-0003-1789-9588","position":6,"is_corresponding":false},{"id":955247,"name":"Barbara Czakó","orcid":"0000-0002-3357-7514","position":7,"is_corresponding":false},{"id":616160,"name":"Jason B. Cross","orcid":"0000-0002-0919-2833","position":8,"is_corresponding":false},{"id":646134,"name":"Philip Jones","orcid":"0000-0001-6149-8293","position":9,"is_corresponding":false},{"id":252487,"name":"Burton F. Dickey","orcid":"0000-0002-4780-1847","position":10,"is_corresponding":false},{"id":57565,"name":"Axel T. Brünger","orcid":"0000-0001-5121-2036","position":11,"is_corresponding":false},{"id":854416,"name":"Ying Lai","orcid":"0000-0001-5347-5155","position":0,"is_corresponding":true}],"reference_count":27,"raw_metadata":null,"created_at":"2026-07-19T00:29:03.247347Z","pmid":"35814209","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":[]}