{"doi":"10.1101/2022.04.14.488403","title":"Cooperative <i>cis</i> -interactions between ectodomains of TCRαβ CD3 subunits enable mechanotransduction","abstract":"ABSTRACT TCR signaling poses a mechanical problem: pMHC binding occurs at the TCRαβ ectodomain (ECD) head, whereas ITAM phosphorylation occurs on CD3 cytoplasmic tails. Chemistry cannot bridge this &gt;10 nm gap, requiring the two events to be coupled through the TCR–CD3 interface, thus involving conformational allostery and being regulatable by force. Although weak ECD cis -interactions between TCRαβ and CD3 have been proposed to contribute to this coupling, their kinetics and mechanical competence remain elusive. Here, we quantify TCRαβ–CD3 ECD cis -interactions in a pseudo- cis configuration using two-dimensional binding and single-bond force spectroscopy, finding that TCRαβ–CD3γε and TCRαβ–CD3δε interactions have low affinity and rapid kinetics, yet form catch bonds. Critically, concurrent engagement of CD3γε and CD3δε produces high cis -cooperativity, yielding a stronger and longer-lasting CD3γε–TCRαβ–CD3δε trimolecular catch bond than the sum of the two dimeric bonds, with force-stabilized lifetimes matching those of agonist TCR–pMHC trans -interaction. Molecular dynamics simulations reveal an expanded, cooperative, and asymmetric contact network, making CD3δε more force-responsive and susceptible to conformational change than CD3γε. Interface mutations do not alter force-free affinities but remodel cooperative cis -bond profiles, leading to an inverse correlation with trans -bond profiles and T cell signaling. These results identify cooperative ECD cis -interaction as a mechanically regulatable allosteric coupling element at the TCR–CD3 junction important to antigen recognition and signal initiation.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":306833,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9469,"is_data_producer":false,"deposit_databanks":null,"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":986061,"name":"Aswin Natarajan","orcid":null,"position":1,"is_corresponding":false},{"id":336859,"name":"Zhou Yuan","orcid":"0000-0003-3437-0125","position":2,"is_corresponding":false},{"id":985511,"name":"Chenghao Ge","orcid":"0000-0002-1386-3177","position":3,"is_corresponding":false},{"id":336860,"name":"Stefano Travaglino","orcid":"0000-0002-7362-2527","position":4,"is_corresponding":false},{"id":1001499,"name":"Saikiran Beesam","orcid":null,"position":5,"is_corresponding":false},{"id":7624,"name":"Xiang Li","orcid":"0000-0002-1945-0381","position":6,"is_corresponding":false},{"id":1001500,"name":"Danielle Grazette","orcid":null,"position":7,"is_corresponding":false},{"id":826969,"name":"Michelle Krogsgaard","orcid":"0000-0001-5006-7981","position":8,"is_corresponding":false},{"id":336861,"name":"Cheng Zhu","orcid":"0000-0002-1718-565X","position":9,"is_corresponding":false},{"id":985510,"name":"Peiwen Cong","orcid":"0000-0001-7753-8368","position":0,"is_corresponding":true}],"reference_count":70,"raw_metadata":null,"created_at":"2026-07-19T00:32:52.703737Z","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":[]}