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The virus distinguishes itself by the ability to replicate to high titers and transmit efficiently in a wide variety of hosts in diverse climatic environments. Fortunately, transmission to and between humans is scarce. Yet, if such an event were to occur, it could spark a pandemic as humans are immunologically na\u00efve to H5 viruses. A significant determinant of transmission to and between humans is the ability of the influenza A virus hemagglutinin (HA) protein to shift from an avian-type to a human-type receptor specificity. Here, we demonstrate that a 2016 2.3.4.4e virus HA can convert to human-type receptor binding via a single Q226L mutation, in contrast to a cleavage-modified 2016 2.3.4.4b virus HA. Using glycan arrays, x-ray structural analyses, tissue- and direct glycan binding, we show that L133a\u0394 and 227Q are vital for this phenotype. Thus, whereas the 2.3.4.4e virus HA only needs a single amino acid mutation, the modified 2.3.4.4b HA was not easily converted to human-type receptor specificity.<\/jats:p>","DOI":"10.1101\/2025.01.10.632119","type":"posted-content","created":{"date-parts":[[2025,1,11]],"date-time":"2025-01-11T04:35:18Z","timestamp":1736570118000},"source":"Crossref","is-referenced-by-count":2,"title":["The Q226L mutation can convert a highly pathogenic H5 2.3.4.4e virus to bind human-type receptors"],"prefix":"10.64898","author":[{"given":"Mar\u00eda R\u00edos","family":"Carrasco","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting-Hui","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xueyong","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alba Gabarroca","family":"Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elif","family":"Uslu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruonan","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cindy M.","family":"Spruit","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathilde","family":"Richard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3111-5954","authenticated-orcid":false,"given":"Geert-Jan","family":"Boons","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian A.","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1586-4464","authenticated-orcid":false,"given":"Robert P.","family":"de Vries","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025011304100891000_2025.01.10.632119v1.1","doi-asserted-by":"publisher","DOI":"10.1111\/irv.12412"},{"key":"2025011304100891000_2025.01.10.632119v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2024.05.018"},{"key":"2025011304100891000_2025.01.10.632119v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07849-4"},{"key":"2025011304100891000_2025.01.10.632119v1.4","doi-asserted-by":"crossref","unstructured":"J. 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