{"doi":"10.1016/j.jbc.2021.100311","title":"T cell receptor–dependent S-acylation of ZAP-70 controls activation of T cells","abstract":"ZAP-70 is a tyrosine kinase essential for T cell immune responses. Upon engagement of the T cell receptor (TCR), ZAP-70 is recruited to the specialized plasma membrane domains, becomes activated, and is released to phosphorylate its laterally segregated targets. A shift in ZAP-70 distribution at the plasma membrane is recognized as a critical step in TCR signal transduction and amplification. However, the molecular mechanism supporting stimulation-dependent plasma membrane compartmentalization of ZAP-70 remains poorly understood. In this study, we identified previously uncharacterized lipidation (S-acylation) of ZAP-70 using Acyl-Biotin Exchange assay, a technique that selectively captures S-acylated proteins. We found that this posttranslational modification of ZAP-70 is dispensable for its enzymatic activity. However, the lipidation-deficient mutant of ZAP-70 failed to propagate the TCR pathway suggesting that S-acylation is essential for ZAP-70 interaction with its protein substrates. The kinetics of ZAP-70 S-acylation were consistent with TCR signaling events indicating that agonist-induced S-acylation is a part of the signaling mechanism controlling T cell activation and function. Taken together, our results suggest that TCR-induced S-acylation of ZAP-70 can serve as a critical regulator of T cell-mediated immunity. ZAP-70 is a tyrosine kinase essential for T cell immune responses. Upon engagement of the T cell receptor (TCR), ZAP-70 is recruited to the specialized plasma membrane domains, becomes activated, and is released to phosphorylate its laterally segregated targets. A shift in ZAP-70 distribution at the plasma membrane is recognized as a critical step in TCR signal transduction and amplification. However, the molecular mechanism supporting stimulation-dependent plasma membrane compartmentalization of ZAP-70 remains poorly understood. In this study, we identified previously uncharacterized lipidation (S-acylation) of ZAP-70 using Acyl-Biotin Exchange assay, a technique that selectively captures S-acylated proteins. We found that this posttranslational modification of ZAP-70 is dispensable for its enzymatic activity. However, the lipidation-deficient mutant of ZAP-70 failed to propagate the TCR pathway suggesting that S-acylation is essential for ZAP-70 interaction with its protein substrates. The kinetics of ZAP-70 S-acylation were consistent with TCR signaling events indicating that agonist-induced S-acylation is a part of the signaling mechanism controlling T cell activation and function. Taken together, our results suggest that TCR-induced S-acylation of ZAP-70 can serve as a critical regulator of T cell-mediated immunity. ZAP-70 (“CD3ζ-chain-associated protein kinase 70”) is one of the first proteins activated upon T cell receptor (TCR) engagement by the peptide/major histocompatibility complex on the surface the antigen-presenting cell (1Brownlie R.J. Zamoyska R. T cell receptor signalling networks: Branched, diversified and bounded.Nat. Rev. Immunol. 2013; 13: 257-269Crossref PubMed Scopus (276) Google Scholar). Stimulation of the TCR leads to activation of the Src-family kinase Lck, which then phosphorylates immunoreceptor tyrosine-based activation motifs (ITAMs) of TCR-associated CD3ζ-chain (1Brownlie R.J. Zamoyska R. T cell receptor signalling networks: Branched, diversified and bounded.Nat. Rev. Immunol. 2013; 13: 257-269Crossref PubMed Scopus (276) Google Scholar, 2Salmond R.J. Filby A. Qureshi I. Caserta S. Zamoyska R. T-cell receptor proximal signaling via the Src-family kinases, Lck and Fyn, influences T-cell activation, differentiation, and tolerance.Immunol. Rev. 2009; 228: 9-22Crossref PubMed Scopus (245) Google Scholar). Double phosphorylation of ITAMs creates a high-affinity docking site for Src homology 2 (SH2) domains of ZAP-70 resulting in its recruitment to the TCR–CD3 complex. Binding to ITAMs triggers conformational changes that make ZAP-70 more accessible to phosphorylation by Lck and subs","journal":"Journal of Biological Chemistry","year":2021,"id":168331,"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":30,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9481,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":567355,"name":"Shayahati Bieerkehazhi","orcid":"0000-0001-9520-4118","position":1,"is_corresponding":false},{"id":497488,"name":"Ying Fan","orcid":"0000-0002-2532-9254","position":2,"is_corresponding":false},{"id":504325,"name":"Askar M. Akimzhanov","orcid":"0000-0002-5592-8215","position":3,"is_corresponding":false},{"id":504115,"name":"Ritika Tewari","orcid":null,"position":0,"is_corresponding":true}],"reference_count":46,"raw_metadata":null,"created_at":"2026-07-18T23:46:02.603031Z","pmid":"33482200","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":[]}