The physicochemical design, structure and permeability of the JCVI-syn3A minimal cell membrane

The physicochemical design, structure and permeability of the JCVI-syn3A minimal cell membrane | 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 Research Article The physicochemical design, structure and permeability of the JCVI-syn3A minimal cell membrane Caterina Presutti, Mert Bozoflu, Joanna Juhaniewicz-Dębińska, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8843141/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 Minimal cells offer a platform to uncover the fundamental physicochemical principles of cellular life. While genomes, proteomes and metabolic networks of cells have been elucidated, the lipidome, which determines the physicochemical identity of the membrane, remains largely unexplored. Yet, the membrane is a central determinant of cellular viability, regulating solute permeability, mechanical stability, and environmental cues. Here, we investigate membranes that recapitulate the lipid composition of the minimal cell JCVI-syn3A, which is distinguished by an unusually high cholesterol content and simple composition. Combining cryo-electron microscopy, Langmuir monolayer experiments, permeability assays, and coarse-grained molecular dynamics simulations, we demonstrate that cholesterol and sphingomyelin act as dominant condensing agents that stabilize the membrane while preserving an ordered yet fluid state. These lipids enhance lipid packing and acyl-chain order, whereas cardiolipin, POPC, and DOPG counterbalance condensation by promoting fluidity and compressibility. Cholesterol–sphingomyelin interactions emerge as a key thermodynamic driver that fine-tunes membrane order, fluidity, and permeability. Remarkably, membranes containing up to 60 mol% of cholesterol remain permeable to water and physiologically relevant osmolytes at rates compatible with growth of JCVI-syn3A. Together, our results define the physicochemical principles underlying minimal cell membranes and reveal how lipid composition enables passive permeation while maintaining membrane integrity. Biophysics physicochemical properties of membranes minimal cell JCVI-syn3A membrane fluidity and permeability Langmuir monolayers molecular dynamics simulations Full Text Additional Declarations The authors declare no competing interests. Supplementary Files SupportingInformationMB3SJM2FINAL.pdf Supplementary Information: The physicochemical design, structure and permeability of the JCVI-syn3A minimal cell membrane 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-8843141","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":589109051,"identity":"3454c759-9ae1-4f56-b01f-2b57dcdf995a","order_by":0,"name":"Caterina Presutti","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Caterina","middleName":"","lastName":"Presutti","suffix":""},{"id":589109052,"identity":"dbc4af61-dc1e-41aa-92bc-321bc4300b2c","order_by":1,"name":"Mert Bozoflu","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Mert","middleName":"","lastName":"Bozoflu","suffix":""},{"id":589109053,"identity":"3c727a7f-7790-494f-9608-4f20e4b7fb21","order_by":2,"name":"Joanna Juhaniewicz-Dębińska","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Joanna","middleName":"","lastName":"Juhaniewicz-Dębińska","suffix":""},{"id":589109054,"identity":"bd45e1af-2fc5-46df-939f-bf6b963c99f2","order_by":3,"name":"Lauren A. 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