{"doi":"10.1186/s12964-025-02435-1","title":"2-chlorofatty acid modification of neutrophil proteins: identification, localization and role in NETosis","abstract":"We previously demonstrated neutrophil MPO derived HOCl targets the vinyl ether bond of plasmalogens resulting in the Liberation of 2-chlorofatty aldehydes (2-ClFALDs) and their oxidation products, 2-chlorofatty acids (2-ClFAs), which elicit neutrophil extracellular trap (NET) formation. In this study, the click chemistry analog of 2-chlorohexadecanoic acid (2-ClHA) was utilized to identify 127 proteins covalently modified by 2-ClHA in human neutrophils. Bioinformatics revealed that multiple proteins modified by 2-ClHA are related to protein modification and binding as well as metabolite interconversion. Three key proteins involved in NET formation and function were modified by 2-ClHA including peptidyl arginine deiminase 4 (PAD4), neutrophil defensin alpha 3 (DEFA3), and neutrophil collagenase (MMP8). PAD4 activity was shown to be increased by 2-ClFA treatment. Further studies investigated 2-ClFA modified protein localization over time during NET formation. Initially PAD4 and 2-ClFA-modified proteins were extranuclear but over time they both localized to distinct nuclear regions. Following DNA release from neutrophils, 2-ClFA-modified proteins were found throughout the neutrophil and DNA strands. In summary, multiple neutrophil proteins are modified by 2-ClHA, including PAD4. 2-ClHA modification and activation of PAD4 is suggested as a key component of 2-ClHA elicited NET formation. • One hundred and twenty-seven neutrophil proteins are covalently modified by 2-ClFA. • 2-ClFA covalently modified human neutrophil proteins including neutrophil defensin alpha 3 (DEFA3), neutrophil collagenase (MMP8) and peptidyl arginine deiminase 4 (PAD4). • Co-localization of DEFA3 and PAD4 with 2-ClFA is maintained over time during NET formation. • Both 2-ClFA and PAD4 colocalize to the nucleus prior to the release of DNA strands during NET formation. • 2-ClFA activates PAD4.","journal":"Cell Communication and Signaling","year":2025,"id":547999,"datarank":0.10397207708399181,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.0,"self_endowment_contribution":0.10397207708399181,"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":0.9638,"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":901090,"name":"Reagan M. McGuffee","orcid":"0000-0001-6516-6413","position":1,"is_corresponding":false},{"id":329406,"name":"Rakesh P. Patel","orcid":"0000-0002-1526-4303","position":2,"is_corresponding":false},{"id":380621,"name":"David A. Ford","orcid":"0000-0002-0029-1560","position":3,"is_corresponding":false},{"id":957904,"name":"Haley Carlson","orcid":"0000-0001-6445-7948","position":0,"is_corresponding":true}],"reference_count":54,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:53:53.962932Z","pmid":"41146178","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":[]}