Conformational landscape of integrin α4β1 and its recognition of VCAM and conformation-stabilizing small molecules

Conformational landscape of integrin α4β1 and its recognition of VCAM and conformation-stabilizing small molecules | 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 Conformational landscape of integrin α4β1 and its recognition of VCAM and conformation-stabilizing small molecules Timothy Springer, Uriel López-Sánchez, Meghan Monroy, Qi Qiao, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8928859/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Integrin α4β1 recognition of vascular cell adhesion molecule (VCAM) is essential for leukocyte adhesion, migration, and signaling, yet is structurally unresolved. Here, we report cryo-electron microscopy (cryo-EM) structures of human α4β1 in apo closed conformations and in an open conformation bound to conformation-specific antibody fragment 12G10, alone or together with VCAM domains 1-3 or 4-6. The homologous D1 and D4 domains recognize α4β1 with comparable affinity, inserting lengthwise between the α4 propeller and β1 βI domains. The VCAM loop that binds the integrin metal ion-dependent adhesion site (MIDAS) flips its backbone and Asp sidechain to coordinate the MIDAS Mg2+ ion. Crystal structures of α4β1 with small-molecule stabilizers of closed or open states reveal a novel closure-stabilizing motif and diverse binding modes, including to α/β interfaces outside the VCAM binding pocket. We define the mechanism of α4β1 activation and ligand recognition, providing a framework for the rational design of immunomodulatory therapeutics. Biological sciences/Structural biology Biological sciences/Chemical biology/Pharmacology Biological sciences/Cell biology Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryInformation.pdf Supplementary Information Cite Share Download PDF Status: Under Review 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-8928859","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":604645974,"identity":"df90e43e-9030-4312-9a55-bf55b0050335","order_by":0,"name":"Timothy 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