Multimodal binding of collybistin controls gephyrin filament formation in synaptic clustering

Multimodal binding of collybistin controls gephyrin filament formation in synaptic clustering | 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 Multimodal binding of collybistin controls gephyrin filament formation in synaptic clustering Guenter Schwarz, Nele Burdina, Filip Liebsch, Arthur Macha, Monika Gunkel, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5890130/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 The multifunctional protein gephyrin clusters glycine and GABA type A receptors at inhibitory postsynapses by oligomerization of its G- and E-domain. Collybistin, a gephyrin-interacting nucleotide exchange factor, mediates gephyrin recruitment and clustering at specific GABAergic postsynapses via phosphoinositide interaction. Using single-particle cryo-electron microscopy, we uncovered the structural basis of collybistin-regulated gephyrin clustering, which controls gephyrin oligomerization in two ways: While collybistin inhibited E-domain dimerization-dependent filament formation of gephyrin, the presence of phosphoinositides promoted the formation of stable gephyrin-collybistin complexes that underwent E-domain dimerization-dependent filament assembly. Within these complexes, collybistin binds at distinct positions, with different stoichiometries and conformations, either promoting or restricting filament assembly and thus regulating filament formation and elongation. Our study highlights the critical role of E-domain dimerization-dependent filament formation for postsynaptic clustering and demonstrates its tight regulation by collybistin, driving specific oligomerization at phosphoinositide-enriched GABAergic synapses. Biological sciences/Biochemistry/Structural biology/Electron microscopy/Cryoelectron microscopy Biological sciences/Biochemistry/Proteins/Membrane proteins Full Text Additional Declarations There is NO Competing Interest. Table 1 is available in the Supplementary Files section. Supplementary Files 20250122GephCBFilamentSupplementaryInformation.pdf Supplemental Information 20250122GephCBFilamentTable1.doc Table 1 20250122GephCBFilamentExtendedData.pdf Extended data figures 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-5890130","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":411012948,"identity":"e99010ed-9ae8-473c-a9d7-76b66d4cf8e8","order_by":0,"name":"Guenter 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Collybistin, a gephyrin-interacting nucleotide exchange factor, mediates gephyrin recruitment and clustering at specific GABAergic postsynapses via phosphoinositide interaction. Using single-particle cryo-electron microscopy, we uncovered the structural basis of collybistin-regulated gephyrin clustering, which controls gephyrin oligomerization in two ways: While collybistin inhibited E-domain dimerization-dependent filament formation of gephyrin, the presence of phosphoinositides promoted the formation of stable gephyrin-collybistin complexes that underwent E-domain dimerization-dependent filament assembly. Within these complexes, collybistin binds at distinct positions, with different stoichiometries and conformations, either promoting or restricting filament assembly and thus regulating filament formation and elongation. Our study highlights the critical role of E-domain dimerization-dependent filament formation for postsynaptic clustering and demonstrates its tight regulation by collybistin, driving specific oligomerization at phosphoinositide-enriched GABAergic synapses.","manuscriptTitle":"Multimodal binding of collybistin controls gephyrin filament formation in synaptic clustering","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-13 05:44:47","doi":"10.21203/rs.3.rs-5890130/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"b57353fd-ba12-4253-aed9-56b7c00207fa","owner":[],"postedDate":"March 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":43820874,"name":"Biological sciences/Biochemistry/Structural biology/Electron microscopy/Cryoelectron microscopy"},{"id":43820875,"name":"Biological sciences/Biochemistry/Proteins/Membrane proteins"}],"tags":[],"updatedAt":"2025-03-13T05:44:48+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-13 05:44:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5890130","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5890130","identity":"rs-5890130","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}