{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T15:58:21Z","timestamp":1786118301150,"version":"build-2736575974"},"posted":{"date-parts":[[2026,8,4]]},"group-title":"Microbiology","reference-count":72,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["1DP2GM146253"],"award-info":[{"award-number":["1DP2GM146253"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R15GM152959"],"award-info":[{"award-number":["R15GM152959"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"award":["P30GM133893"],"award-info":[{"award-number":["P30GM133893"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"name":"NSF","award":["2205006"],"award-info":[{"award-number":["2205006"]}]},{"name":"Army Reseach Office","award":["W911NF-25-1-0084"],"award-info":[{"award-number":["W911NF-25-1-0084"]}]},{"DOI":"10.13039\/100000879","name":"Alfred P. 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To be effective, a biofilm must anchor to foreign surfaces and retain sufficient stiffness, in the environment or in a host. However, how this matrix self-organizes to support biofilm formation remains a mystery at the molecular level. The human pathogen\n                  <jats:italic>Vibrio cholerae<\/jats:italic>\n                  produces biofilms primarily composed of an exopolysaccharide called VPS (\n                  <jats:underline>V<\/jats:underline>\n                  ibrio\n                  <jats:underline>p<\/jats:underline>\n                  oly\n                  <jats:underline>s<\/jats:underline>\n                  accharide), consisting of an unusually-modified repeating tetrasaccharide core unit. VPS engages with two secreted adhesion proteins, Bap1 and RbmC, which adhere the biofilm to abiotic and biotic surfaces, and serve to strengthen the biofilm by interacting with VPS using a conserved \u03b2-propeller. To pinpoint the interaction between the adhesins and purified segments of VPS, we determined the \u223c1.6 \u00c5 X-ray crystal structure of Bap1 bound to fragmented VPS and used the structure to carry out molecular dynamics simulations. The structure revealed a single binding site consisting of one tetrasaccharide unit involving an induced magnesium binding site. Unexpectedly, the tetrasaccharide adopted a bent state caused by a rotation of the glycosidic bond between the central two monosaccharide units. Using a combination of mutagenesis, light scattering, and\n                  <jats:italic>in situ<\/jats:italic>\n                  fluorescent microscopy, we demonstrate that Bap1 not only facilitates biofilm adhesion, but is also required for proper VPS organization, through the identified binding pocket. Our structure reveals for the first time the interaction between a biofilm exopolysaccharide and matrix protein, as well as insights into conformational changes of exopolysaccharide induced by this binding. Our findings provide a generalizable approach for studying the biophysical and biochemical properties of carbohydrate-dependent biofilm assembly, which may lead to new ways to treat disease caused by biofilm-forming bacterial pathogens by disrupting the exopolysaccharide-protein interactions.\n                <\/jats:p>","DOI":"10.64898\/2026.08.03.742583","type":"posted-content","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T04:52:14Z","timestamp":1785905534000},"source":"Crossref","is-referenced-by-count":0,"title":["Structure of a key adhesin-exopolysaccharide interaction provides insights into matrix assembly in\n                  <i>Vibrio cholerae<\/i>\n                  biofilms"],"prefix":"10.64898","author":[{"given":"Alexander J.","family":"Hinbest","sequence":"first","affiliation":[{"name":"Department of Molecular Biology and Biochemistry, Molecular Biophysics Program, Wesleyan University, Middletown, CT, 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