Structural basis of collagen glucosyltransferase function and its serendipitous role in kojibiose synthesis

Structural basis of collagen glucosyltransferase function and its serendipitous role in kojibiose synthesis | 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 Structural basis of collagen glucosyltransferase function and its serendipitous role in kojibiose synthesis Hou-Fu Guo, Jeong Seon Kim, Zhenhang Chen, Sara Espinosa Garcia, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5850681/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Jul, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Collagen glucosyltransferases catalyze a unique type of collagen glucosylation that is critical for biological processes and disease mechanisms. However, the structural regulation of collagen glucosyltransferases remains poorly understood. Here, we report the crystal structures of a mimiviral collagen glucosyltransferase in its apo form and in complexes with uridine diphosphate (UDP) and the disaccharide product. Our findings reveal that the enzyme functions as a homodimer, stabilized by a loop from one subunit locking into a cleft on the opposite subunit. This dimerization enables UDP-glucose binding cooperativity and enzymatic activity, a property conserved in the human homolog. Further structural analyses suggest an induced fit model for UDP interaction, mediated by Lysine 222. The dimerization also forms an extended cleft flanked by two active sites, which likely facilitates collagen recognition. Unexpectedly, we discovered that the mimiviral collagen glucosyltransferase can also synthesize the prebiotic disaccharide kojibiose. An elongated pocket adjacent to the UDP-binding site allows the enzyme to use UDP-glucose as the sugar donor and glucose as the acceptor for kojibiose production. Enzymatic activity assays confirmed the enzyme's novel kojibiose synthesis activity in vitro and in vivo. These structural insights not only inform glucosyltransferase function but also open new avenues for biomedicine. Biological sciences/Biochemistry/Structural biology/X-ray crystallography Biological sciences/Biochemistry/Glycobiology lysyl hydroxylase glycosylation lysyl post-translational modifications DNA viruses Full Text Additional Declarations There is NO Competing Interest. Supplementary Files R699SI9DYT9DZS9E9220250108.docx Crystallographic tables D1000287937valreportfullP1.pdf Crystal structure validation report 1 D1000287938valreportfullP1.pdf Crystal structure validation report 2 D1000287889valreportfullP1.pdf Crystal structure validation report 3 Cite Share Download PDF Status: Published Journal Publication published 21 Jul, 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. 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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-5850681","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":407907060,"identity":"6f4e5d79-a46e-4317-925e-64f0293b334c","order_by":0,"name":"Hou-Fu 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An elongated pocket adjacent to the UDP-binding site allows the enzyme to use UDP-glucose as the sugar donor and glucose as the acceptor for kojibiose production. Enzymatic activity assays confirmed the enzyme's novel kojibiose synthesis activity in vitro and in vivo. 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