{"doi":"10.1101/2025.11.02.686167","title":"Signaling bias of the protease-activated receptor-1 is dictated by distinct GRK5 and β-arrestin-2 determinants","abstract":"G protein-coupled receptors (GPCRs) exhibit signaling bias, or preferential activation of heterotrimeric G proteins versus GPCR receptor kinase (GRK)-mediated β-arrestin signaling. The protease-activated receptor-1 (PAR1) GPCR activates both G protein and β-arrestin in response to thrombin, but only β-arrestin in response to activated protein C (APC). Thrombin-activated PAR1-G protein signaling is desensitized by β-arrestin-1, whereas APC-activated PAR1 signaling is propagated by β-arrestin-2. The mechanisms underlying PAR1 biased signaling are not known. Here, using computational modeling and cell biology studies, we reveal the molecular basis of signaling by thrombin- and APC-activated PAR1. Although both thrombin- and APC-induced PAR1 signaling are regulated by the same GRK isoform, GRK5, the two types of signaling are differentially dependent on GRK5 membrane anchoring, PAR1 C-terminal phosphorylation, and the binding mode of β-arrestin-2. These differences translate into distinct β-arrestin-2 conformations and define the cytoprotective signaling signature by APC which contrasts with thrombin inflammatory signaling.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":556869,"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":0.9589,"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":699180,"name":"Lennis B. Orduña-Castillo","orcid":"0000-0001-7013-861X","position":1,"is_corresponding":false},{"id":98053,"name":"Carolyne Bardeleben","orcid":null,"position":2,"is_corresponding":false},{"id":1002245,"name":"Huaping Qin","orcid":null,"position":3,"is_corresponding":false},{"id":465531,"name":"Ying Lin","orcid":"0000-0002-5619-1104","position":4,"is_corresponding":false},{"id":475851,"name":"Cierra A. Birch","orcid":null,"position":5,"is_corresponding":false},{"id":354098,"name":"Irina Kufareva","orcid":"0000-0001-9083-7039","position":6,"is_corresponding":false},{"id":247561,"name":"JoAnn Trejo","orcid":"0000-0003-4405-6228","position":7,"is_corresponding":false},{"id":910577,"name":"Monica L. Gonzalez Ramirez","orcid":null,"position":0,"is_corresponding":true}],"reference_count":61,"raw_metadata":null,"created_at":"2026-07-19T02:55:13.130091Z","pmid":"41279546","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":[]}