{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T13:25:18Z","timestamp":1785936318103,"version":"3.56.0"},"posted":{"date-parts":[[2025,12,30]]},"group-title":"Microbiology","reference-count":50,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T00:00:00Z","timestamp":1767052800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R01AI189907"],"award-info":[{"award-number":["R01AI189907"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R01AI087946"],"award-info":[{"award-number":["R01AI087946"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R01AI132818"],"award-info":[{"award-number":["R01AI132818"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["S10OD023603"],"award-info":[{"award-number":["S10OD023603"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,12,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The bacterial flagellum is a complex nanomachine essential for motility, colonization, and invasion in diverse species.\n                  <jats:italic>Helicobacter pylori<\/jats:italic>\n                  has evolved elaborate sheathed flagella that enable migration through the highly viscous gastric mucus layer to reach its colonization niche on the gastric epithelium, yet the molecular basis for these unique adaptations has remained elusive. Here, we use\n                  <jats:italic>in-situ<\/jats:italic>\n                  single particle cryo-electron microscopy to determine near-atomic structures of the flagellar filament within the membranous sheath of\n                  <jats:italic>H. pylori<\/jats:italic>\n                  . The major flagellin FlaA constitutes the bulk of the filament, whereas the minor flagellin FlaB contributes critically to the hook-proximal region. Both FlaA and FlaB form a conserved core surrounded by variable surface-exposed domains. Our structures further reveal that pseudaminic-acid glycans decorate these domains, where they mediate inter- and intra-subunit contacts that stabilize the filament and confer a negatively charged surface. Together, these findings support a model in which the filament rotates independently of the membranous sheath to drive\n                  <jats:italic>H. pylori<\/jats:italic>\n                  motility and provide a molecular framework for understanding how the sheathed flagellum enables colonization and persistence within the gastric niche.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance Statement<\/jats:title>\n                  <jats:p>\n                    We present the first\n                    <jats:italic>in-situ<\/jats:italic>\n                    near-atomic structure of the sheathed flagellar filament in\n                    <jats:italic>Helicobacter pylori<\/jats:italic>\n                    , revealing distinctive adaptations that underpin the pathogen\u2019s unique motility and persistent infection. Our\n                    <jats:italic>in-situ<\/jats:italic>\n                    structures show that the two flagellins, FlaA and FlaB, assemble into an extended and exceptionally stable filament through an extensive hydrogen-bonding network. Pseudaminic acid glycans decorate the surface-exposed domains, where they stabilize inter-subunit packing and render the surface negatively charged and hydrophilic. These findings, which provide insight into the assembly of the flagellar filament and its relationship to the surrounding sheath, provide a structural framework for developing strategies to disrupt\n                    <jats:italic>H. pylori<\/jats:italic>\n                    motility and infection.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2025.12.30.697013","type":"posted-content","created":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T23:05:12Z","timestamp":1767135912000},"source":"Crossref","is-referenced-by-count":2,"title":["Assembly and glycosylation of\n                  <i>Helicobacter pylori<\/i>\n                  sheathed flagella"],"prefix":"10.64898","author":[{"given":"Rajeev","family":"Kumar","sequence":"first","affiliation":[{"name":"Department of Microbial Pathogenesis, Yale School of Medicine, New Haven, CT USA"},{"name":"Microbial Sciences Institute, Yale University, West Haven, CT 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