{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,31]],"date-time":"2026-10-31T05:34:14Z","timestamp":1793424854061,"version":"4.5.0"},"posted":{"date-parts":[[2026,6,1]]},"group-title":"Biophysics","reference-count":67,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T00:00:00Z","timestamp":1780272000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R35GM124779"],"award-info":[{"award-number":["R35GM124779"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["T32EY023211"],"award-info":[{"award-number":["T32EY023211"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R24GM154185"],"award-info":[{"award-number":["R24GM154185"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["S10OD034224"],"award-info":[{"award-number":["S10OD034224"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,6,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>Gap junction channels enable direct electrical and metabolic exchange between adjacent cells and tissues, where they organize into dense plaques containing tens to thousands of channels. Although plaque formation is known to modulate junctional conductance, the structural basis for channel\u2013channel organization within a membrane environment remains poorly defined. Here, we reconstitute native lens connexin-46\/50 (Cx46\/50) gap junction channels into MSP-based lipid nanodiscs that incorporate multiple channels to create miniature plaque-like complexes suitable for single-particle cryo-electron microscopy (cryo-EM). We determine high-resolution structures of dual-channel assemblies in two distinct configurations and find no ordered protein\u2013protein contacts across the interface. Instead, the inter-channel space is occupied by ordered lipid density, indicating that channel packing in these assemblies is lipid mediated. These channel\u2013channel interfaces stabilize discrete lipid populations, including an interstitial lipid intercalated between subunits and positioned near the N-terminal gating domain, suggesting a route by which channel organization could promote lipid occupancy near the pore. Additionally, we leverage this dataset to refine the Cx46\/50 single-channel structure to 1.8 \u00c5 resolution, revealing exceptional chemical detail of the pore-lining landscape in the stabilized open-state. Together, these results define principles of lipid-mediated multi-channel organization and suggest how plaque-like packing may tune gap junction function through specific lipid interactions.<\/jats:p>","DOI":"10.64898\/2026.05.28.728492","type":"posted-content","created":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T17:55:15Z","timestamp":1780336515000},"source":"Crossref","is-referenced-by-count":1,"title":["Lipids intercalate and mediate multi-channel assemblies of connexin-46\/50 gap junctions"],"prefix":"10.64898","author":[{"given":"Connor S.","family":"Garrels","sequence":"first","affiliation":[{"name":"Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR 97239, USA"},{"name":"Vollum Institute, Oregon Health & Science University, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janette B.","family":"Myers","sequence":"additional","affiliation":[{"name":"Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR 97239, USA"},{"name":"Vollum Institute, Oregon Health & Science University, Portland, OR 97239, USA"},{"name":"Pacific Northwest Cryo-EM Center, Oregon Health & Science University, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samson A.","family":"Souza","sequence":"additional","affiliation":[{"name":"Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR 97239, USA"},{"name":"Vollum Institute, Oregon Health & Science University, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua M.","family":"Jarodsky","sequence":"additional","affiliation":[{"name":"Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR 97239, USA"},{"name":"Vollum Institute, Oregon Health & Science University, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5276-7690","authenticated-orcid":false,"given":"Steve L.","family":"Reichow","sequence":"additional","affiliation":[{"name":"Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR 97239, USA"},{"name":"Vollum Institute, Oregon Health & Science University, Portland, OR 97239, USA"},{"name":"Pacific Northwest Cryo-EM Center, Oregon Health & Science University, Portland, OR 97239, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026060402350551000_2026.05.28.728492v1.1","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.0000251936.26233.0d"},{"key":"2026060402350551000_2026.05.28.728492v1.2","doi-asserted-by":"publisher","DOI":"10.1002\/cphy.c110051"},{"key":"2026060402350551000_2026.05.28.728492v1.3","doi-asserted-by":"crossref","unstructured":"Delmar, M. et al. 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