{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,31]],"date-time":"2026-10-31T03:27:03Z","timestamp":1793417223776,"version":"4.5.0"},"reference-count":83,"publisher":"National Academy of Sciences","issue":"16","license":[{"start":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T00:00:00Z","timestamp":1744675200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100019180","name":"EC | ERC | HORIZON EUROPE European Research Council","doi-asserted-by":"publisher","award":["802780)"],"award-info":[{"award-number":["802780)"]}],"id":[{"id":"10.13039\/100019180","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009578","name":"Mizutani Foundation for Glycoscience","doi-asserted-by":"publisher","award":["2023"],"award-info":[{"award-number":["2023"]}],"id":[{"id":"10.13039\/100009578","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016305","name":"Animal Health and Welfare ERA-Net","doi-asserted-by":"publisher","award":["862605"],"award-info":[{"award-number":["862605"]}],"id":[{"id":"10.13039\/100016305","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100024870","name":"NWO | Exacte en Natuurwetenschappen","doi-asserted-by":"publisher","award":["OCENW.M20.106"],"award-info":[{"award-number":["OCENW.M20.106"]}],"id":[{"id":"10.13039\/501100024870","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003246","name":"Nederlandse Organisatie voor Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["TOP-PUNT 718.015.003"],"award-info":[{"award-number":["TOP-PUNT 718.015.003"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100015691","name":"HHS | NIH | NIAID | Division of Microbiology and Infectious Diseases","doi-asserted-by":"publisher","award":["75N93021C00015 \/ PENN CEIRR"],"award-info":[{"award-number":["75N93021C00015 \/ PENN CEIRR"]}],"id":[{"id":"10.13039\/100015691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,4,22]]},"abstract":"<jats:p>H5Nx viruses continue to wreak havoc in avian and mammalian species worldwide. 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 2016 2.3.4.4b HA was not easily converted to human-type receptor specificity.<\/jats:p>","DOI":"10.1073\/pnas.2419800122","type":"journal-article","created":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T11:51:21Z","timestamp":1744717881000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":17,"title":["The Q226L mutation can convert a highly pathogenic H5 2.3.4.4e virus to bind human-type receptors"],"prefix":"10.1073","volume":"122","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7000-2018","authenticated-orcid":false,"given":"Mar\u00eda","family":"R\u00edos Carrasco","sequence":"first","affiliation":[{"name":"Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0937-9972","authenticated-orcid":false,"given":"Ting-Hui","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Integrative Structural and Computational Biology, The Scripps Research Institute","place":["La Jolla"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6021-3740","authenticated-orcid":false,"given":"Xueyong","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Integrative Structural and Computational Biology, The Scripps Research Institute","place":["La Jolla"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4266-3712","authenticated-orcid":false,"given":"Alba","family":"Gabarroca Garc\u00eda","sequence":"additional","affiliation":[{"name":"Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3678-1893","authenticated-orcid":false,"given":"Elif","family":"Uslu","sequence":"additional","affiliation":[{"name":"Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruonan","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cindy M.","family":"Spruit","sequence":"additional","affiliation":[{"name":"Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0240-9312","authenticated-orcid":false,"given":"Mathilde","family":"Richard","sequence":"additional","affiliation":[{"name":"Department of Viroscience, Erasmus University Medical Center","place":["Rotterdam, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3111-5954","authenticated-orcid":false,"given":"Geert-Jan","family":"Boons","sequence":"additional","affiliation":[{"name":"Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University","place":["Utrecht, The Netherlands"]},{"name":"Complex Carbohydrate Research Center, University of Georgia","place":["Athens"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6469-2419","authenticated-orcid":false,"given":"Ian A.","family":"Wilson","sequence":"additional","affiliation":[{"name":"Department of Integrative Structural and Computational Biology, The Scripps Research Institute","place":["La Jolla"]},{"name":"Skaggs Institute for Chemical Biology, The Scripps Research Institute","place":["La Jolla"]}],"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":[{"name":"Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2025,4,15]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1111\/irv.12412"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2024.05.018"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07849-4"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2022.2155072"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1999.9820"},{"key":"e_1_3_4_6_2","unstructured":"WHO & GIP Cumulative number of confirmed human cases for avian influenza A(H5N1) reported to WHO 2003\u20132024. https:\/\/cdn.who.int\/media\/docs\/default-source\/influenza\/h5n1-human-case-cumulative-table\/2024_feb_tableh5n1.pdf?sfvrsn=bccd8c23_1&download=true. Accessed 26 February 2024."},{"key":"e_1_3_4_7_2","unstructured":"CDC \u201cTechnical report: June 2024 Highly pathogenic Avian influenza A(H5N1) viruses\u201d (Tech. Rep. June 2024 US Centers for Disease Control and Prevention 2024)."},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2013.02.015"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/304076a0"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.9.7626-7631.1998"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1136212"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.adt0180"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02690-13"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02016-17"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2011.10.006"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1213362"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature10831"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2015.10.027"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2014.03.008"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature12144"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1236787"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006390"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2017.03.054"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2021.1872355"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2711.211225"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2022.11.008"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES.2023.28.3.2300001"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2904.221538"},{"key":"e_1_3_4_29_2","doi-asserted-by":"crossref","unstructured":"P. Chopra Receptor binding specificity of a bovine A(H5N1) influenza virus. bioRxiv [Preprint] (2024). https:\/\/doi.org\/10.1101\/2024.07.30.605893 (Accessed 30 July 2024).","DOI":"10.1101\/2024.07.30.605893"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-54934-3"},{"key":"e_1_3_4_31_2","doi-asserted-by":"crossref","unstructured":"J. J. S. Santos Bovine H5N1 influenza virus binds poorly to human-type sialic acid receptors. bioRxiv [Preprint] (2024). https:\/\/doi.org\/10.1101\/2024.08.01.606177 (Accessed 1 August 2024).","DOI":"10.1101\/2024.08.01.606177"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3002916"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.3390\/v12121439"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00195-20"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1292\/jvms.21-0459"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2018.12.014"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-05961-x"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003223"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-08246-7"},{"key":"e_1_3_4_40_2","doi-asserted-by":"crossref","unstructured":"L. Bauer A 2022 avian H5N1 influenza A virus from clade 2.3.4.4b attaches to and replicates better in human respiratory epithelium than a 2005 H5N1 virus from clade 2.3.2.1. bioRxiv [Preprint] (2024). https:\/\/doi.org\/10.1101\/2024.11.27.625596 (Accessed 27 November 2024).","DOI":"10.1101\/2024.11.27.625596"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1375"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2016.11.004"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1243761"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.meegid.2022.105281"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-023-06631-2"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.74.18.8502-8512.2000"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009566"},{"key":"e_1_3_4_48_2","doi-asserted-by":"crossref","unstructured":"R. Liang Epistasis in the receptor binding domain of contemporary H3N2 viruses that reverted to bind sialylated diLacNAc repeats. bioRxiv [Preprint] (2024). https:\/\/doi.org\/10.1101\/2024.11.26.625384 (Accessed 26 November 2024).","DOI":"10.1101\/2024.11.26.625384"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1099\/jgv.0.001672"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02790-15"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00127-16"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1062882"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2112.150809"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.03736-23"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-017-3631-7"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00200-23"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-38415-7"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-69535-5"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0129827"},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07766-6"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-08254-7"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2018.03.018"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.01710-2016"},{"key":"e_1_3_4_64_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-47635-4"},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2302.161072"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-015-2660-3"},{"key":"e_1_3_4_67_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwx026"},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1002\/chem.202401108"},{"key":"e_1_3_4_69_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01178-19"},{"key":"e_1_3_4_70_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01366-21"},{"key":"e_1_3_4_71_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2019.05.048"},{"key":"e_1_3_4_72_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1209"},{"key":"e_1_3_4_73_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2010.03.047"},{"key":"e_1_3_4_74_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01363-19"},{"key":"e_1_3_4_75_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.05112-11"},{"key":"e_1_3_4_76_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(97)76066-X"},{"key":"e_1_3_4_77_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889807021206"},{"key":"e_1_3_4_78_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444910007493"},{"key":"e_1_3_4_79_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909052925"},{"key":"e_1_3_4_80_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909042073"},{"key":"e_1_3_4_81_2","doi-asserted-by":"crossref","unstructured":"T. H. Lin X. Zhu I. A. Wilson Crystal structure of H5 hemagglutinin from the influenza virusA\/black swan\/Akita\/1\/2016 with LSTa. https:\/\/www.rcsb.org\/structure\/9NR2. Deposited 13 March 2025.","DOI":"10.2210\/pdb9dip\/pdb"},{"key":"e_1_3_4_82_2","doi-asserted-by":"crossref","unstructured":"T. H. Lin X. Zhu I. A. Wilson Crystal structure of H5 hemagglutinin Q226L mutant from the influenza virusA\/black swan\/Akita\/1\/2016 with LSTc. https:\/\/www.rcsb.org\/structure\/9NR5. Deposited 13 March 2025.","DOI":"10.2210\/pdb9dio\/pdb"},{"key":"e_1_3_4_83_2","doi-asserted-by":"crossref","unstructured":"T. H. Lin X. Zhu I. A. Wilson Crystal structure of H5 hemagglutinin Q226L mutant from the influenza virusA\/duck\/France\/1611008h\/2016 with LSTc. https:\/\/www.rcsb.org\/structure\/9NRB. Deposited 14 March 2025.","DOI":"10.2210\/pdb9dio\/pdb"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2419800122","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T11:51:34Z","timestamp":1744717894000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2419800122"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,15]]},"references-count":83,"journal-issue":{"issue":"16","published-print":{"date-parts":[[2025,4,22]]}},"alternative-id":["10.1073\/pnas.2419800122"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2419800122","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2025.01.10.632119","asserted-by":"object"}]},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,15]]},"assertion":[{"value":"2024-10-01","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-02-24","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-15","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2419800122"}}