Distinct GRK2 Recruitment Mechanisms Differentially Shape GPCR Signaling Bias

Distinct GRK2 Recruitment Mechanisms Differentially Shape GPCR Signaling Bias | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Distinct GRK2 Recruitment Mechanisms Differentially Shape GPCR Signaling Bias Prashant Donthamsetti, Edwin Matthiessen, Madeline Newlin, Kole Martin, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8865860/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Biased GPCR agonists that selectively engage G proteins or arrestins show therapeutic promise, but their development has been challenging. Key to understanding biased signaling is GRK2, which phosphorylates many GPCRs and facilitates arrestin recruitment. Canonically, GRK2 binds GPCRs only after being recruited to the plasma membrane by Gβγ, implying that arrestin recruitment requires prior G protein activation and challenging the feasibility of developing highly biased agonists. However, emerging evidence suggests that certain GPCRs, including dopamine D2 receptor, can bind GRK2 independently of Gβγ. Here, we combined a robust GRK2 recruitment assay and functional analyses to show that Gβγ-dependent and -independent GRK2 recruitment have distinct dynamics, signaling consequences, and impacts on bias. We further demonstrate that Gβγ-independent GRK2 recruitment facilitates arrestin recruitment to diverse GPCRs, revising the canonical model of GRK2 function. These findings highlight the need to account for distinct GRK2 recruitment mechanisms in the design of biased GPCR agonists. Biological sciences/Drug discovery/Pharmacology/Receptor pharmacology Biological sciences/Chemical biology/Pharmacology/Receptor pharmacology Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryFigures.pdf Supplementary Figures RS.pdf Reporting Summary Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8865860","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":597136484,"identity":"9c26fe6d-d329-4af9-8eff-b1aa0b1f8d5b","order_by":0,"name":"Prashant 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