Single-molecule tweezers decoding hidden dimerization patterns of membrane proteins within lipid bilayers

Single-molecule tweezers decoding hidden dimerization patterns of membrane proteins within lipid bilayers | 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 Single-molecule tweezers decoding hidden dimerization patterns of membrane proteins within lipid bilayers Duyoung Min, Victor Sadongo, Eojin Kim, Seoyoon Kim, W.C. Bhashini Wijesinghe, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5918517/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 08 Aug, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Dimerization of transmembrane (TM) proteins is an essential biological process within cellular membranes, playing a key role in diverse pathophysiological pathways and serving as a promising therapeutic target. Although often simplified as a two-state transition from freely diffusing monomers to fully formed dimers, the dimerization process after monomer diffusion—the post-diffusion dimerization—is likely more complex due to intricate inter-residue interactions. Here, we introduce a single-molecule tweezer platform to map detailed profiles of the post-diffusion transitions in TM protein dimerization. This approach captures reversible dimerization events of a single TM dimer, revealing hidden intermediate states that emerge following the quiescent phase of monomer diffusion. Profiling the post-diffusion intermediates, kinetics, and energy landscapes—integrated with molecular dynamics simulations—uncovers the dimerization pathway, the effects of residue interactions and lipid bilayers, and the kinetic and energetic contributions of distinct dimerization domains. Furthermore, this platform characterizes selective and localized modulations via peptide binding, underscoring its potential to elucidate the mechanisms of action of TM dimer-targeting drugs at single-molecule resolution. Biological sciences/Biotechnology/Nanobiotechnology/Biosensors Physical sciences/Physics/Biological physics Full Text Additional Declarations There is NO Competing Interest. Supplementary Files MPPISI.pdf Supplementary Information Cite Share Download PDF Status: Published Journal Publication published 08 Aug, 2025 Read the published version in Nature Communications → 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-5918517","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":425613591,"identity":"3903f567-e2bd-472a-add4-0514cfccb2e2","order_by":0,"name":"Duyoung Min","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAx0lEQVRIiWNgGAWjYDCCAyBUwcDABuUbEKnlDEgLMwlaGBjbQCxitfDdyE48XDjvcB4f//kDDD9qGIzNGwhokbyRu+HwzG2Hi9kkkhkYe44xmMkcIKDFAKSFd9vhxDYJoMN4GxhsJAg5DKJlDlAL/2EGxr/Ea2kAamFIZmAG2mJGUIvkmbcbDvMcSwc6LNngsMwxCWOCWviO527+zFNjnTi//+DDh29qbAxnENLCIJCAYB9gYCBoBxDwHyBC0SgYBaNgFIxsAAARGUASWwrNHQAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-2856-8082","institution":"Ulsan National Institute of Science and Technology","correspondingAuthor":true,"prefix":"","firstName":"Duyoung","middleName":"","lastName":"Min","suffix":""},{"id":425613592,"identity":"e51589d2-8db3-4c1e-8182-d0c447322e11","order_by":1,"name":"Victor Sadongo","email":"","orcid":"","institution":"Ulsan National Institute of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Victor","middleName":"","lastName":"Sadongo","suffix":""},{"id":425613593,"identity":"34705447-4059-490d-a41a-9b6f352ec512","order_by":2,"name":"Eojin Kim","email":"","orcid":"","institution":"Ulsan National Institute of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Eojin","middleName":"","lastName":"Kim","suffix":""},{"id":425613594,"identity":"72b9a17e-3d3c-4395-8d0b-3ed75508007a","order_by":3,"name":"Seoyoon Kim","email":"","orcid":"","institution":"Ulsan National Institute of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Seoyoon","middleName":"","lastName":"Kim","suffix":""},{"id":425613595,"identity":"9a3433fe-98e5-4038-a323-003692361f1f","order_by":4,"name":"W.C. 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Although often simplified as a two-state transition from freely diffusing monomers to fully formed dimers, the dimerization process after monomer diffusion—the post-diffusion dimerization—is likely more complex due to intricate inter-residue interactions. Here, we introduce a single-molecule tweezer platform to map detailed profiles of the post-diffusion transitions in TM protein dimerization. This approach captures reversible dimerization events of a single TM dimer, revealing hidden intermediate states that emerge following the quiescent phase of monomer diffusion. Profiling the post-diffusion intermediates, kinetics, and energy landscapes—integrated with molecular dynamics simulations—uncovers the dimerization pathway, the effects of residue interactions and lipid bilayers, and the kinetic and energetic contributions of distinct dimerization domains. 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