pH-Triggered Clustering Regulates β-sheet Activation in Silk Assembly

pH-Triggered Clustering Regulates β-sheet Activation in Silk Assembly | 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 pH-Triggered Clustering Regulates β-sheet Activation in Silk Assembly Cedric Dicko, Juanita Francis, Judith Houston, Andrew Jackson, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7495345/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Jan, 2026 Read the published version in Communications Chemistry → Version 1 posted You are reading this latest preprint version Abstract Silk fibres derive their exceptional properties from hierarchical protein organisation, yet the molecular pathways guiding this evolution remain poorly resolved. During regenerated silk fibroin gelation under biomimetic gradual acidification, we resolve a stepwise assembly pathway comprising nanoscale clustering, growth of domains within clusters, and mesoscale network formation. Time-resolved small-angle neutron scattering performed simultaneously with UV and fluorescence emission (NUrF) identifies unique intermediates and a regulated onset of β-sheet assembly, indicating that fibril formation requires prior compaction and network connectivity. By contrast, methanol-induced gelation bypasses these intermediates, driving rapid aggregation. These findings define the sequence and timing of events that construct silk’s hierarchical architecture without accidental aggregation, showing how pathway selection governs material outcomes. This multiscale resolution achieved by NurF provides a broadly applicable strategy for probing hierarchical assembly in silk and other protein materials. Biological sciences/Biophysics/Molecular biophysics Biological sciences/Structural biology/SAXS Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryInformation.pdf Supplementary Information Cite Share Download PDF Status: Published Journal Publication published 09 Jan, 2026 Read the published version in Communications Chemistry → 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-7495345","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":509224113,"identity":"e31f9f6f-d8a0-4ba6-98bc-0048f71334fa","order_by":0,"name":"Cedric 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