{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T14:43:15Z","timestamp":1787496195806,"version":"build-2736575974"},"reference-count":81,"publisher":"National Academy of Sciences","issue":"44","license":[{"start":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T00:00:00Z","timestamp":1761782400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000038","name":"Canadian Government | Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["RGPIN-2020-07113"],"award-info":[{"award-number":["RGPIN-2020-07113"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000038","name":"Canadian Government | Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["RGPIN-2018-04365"],"award-info":[{"award-number":["RGPIN-2018-04365"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000024","name":"Canadian Government | Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["FPE-194084"],"award-info":[{"award-number":["FPE-194084"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009056","name":"UofA | Canadian Glycomics Network","doi-asserted-by":"publisher","award":["SD-1"],"award-info":[{"award-number":["SD-1"]}],"id":[{"id":"10.13039\/501100009056","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":[[2025,11,4]]},"abstract":"<jats:p>\n                    <jats:sc>l<\/jats:sc>\n                    -Fucose, a 6-deoxy-monosaccharide, is often incorporated into O-antigens in Rhizobia where it serves as an important marker for host recognition. Here, we biochemically and structurally characterize WrtF, an O-antigen polysaccharide\n                    <jats:sc>l<\/jats:sc>\n                    -fucosyltransferase from the plant endosymbiont\n                    <jats:italic toggle=\"yes\">Rhizobium tropici<\/jats:italic>\n                    CIAT 899. We show that WrtF transfers\n                    <jats:sc>l<\/jats:sc>\n                    -fucose from its donor, guanosine 5\u2032-diphospho-\u03b2-\n                    <jats:sc>l<\/jats:sc>\n                    -fucose (GDP-Fuc), to a nonreducing end glucose, creating an \u03b1-(1\u21924) linkage. We determined structures of WrtF at resolutions between 1.45 and 2.3 \u00c5 in the apo state, and in complex with GDP, GDP-Fuc, nonacetylated and acetylated trisaccharide acceptors, and with a tetrasaccharide product. WrtF has an N-terminal glycosyltransferase A (GT-A) fold and a unique C-terminal \u03b1-helical bundle domain. The structures, combined with activity assays, and sequence analysis show that WrtF is unusual in using neither divalent cations nor basic residues to coordinate and stabilize the donor pyrophosphate. In contrast to almost all previously characterized GT-A glycosyltransferases that use \u03b1-linked donors that present the donor monosaccharide axially, GDP-Fuc presents its monosaccharide equatorially. GDP-Fuc binds in a compact conformation with the\n                    <jats:sc>l<\/jats:sc>\n                    -fucose endocyclic oxygen and C6 methyl packed against ribose C5 and C4, respectively. This binding mode paradoxically shields the anomeric carbon from attack; superposition of the donor and acceptor complexes suggests that these substrates would sterically hinder one another and do not appropriately orient to form a Michaelis complex. We therefore propose that the donor\n                    <jats:sc>l<\/jats:sc>\n                    -fucose must reorient in the active site prior to turnover. The preferred occluded donor binding mode may serve to \u201cprescreen\u201d GDP-Fuc from structurally similar candidate donors, while also minimizing counterproductive donor hydrolysis.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2512460122","type":"journal-article","created":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T17:43:07Z","timestamp":1761846187000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":2,"title":["WrtF from\n                    <i>Rhizobium tropici<\/i>\n                    CIAT 899 is a GT-A fold fucosyltransferase that binds its donor nonproductively"],"prefix":"10.1073","volume":"122","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5609-1453","authenticated-orcid":false,"given":"Taylor J. B.","family":"Forrester","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph","place":["Guelph, Canada"]},{"name":"Department of Anatomy and Cell Biology, McGill University","place":["Montr\u00e9al, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4390-6780","authenticated-orcid":false,"given":"Sicheng","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Alberta","place":["Edmonton, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8331-8211","authenticated-orcid":false,"given":"Todd L.","family":"Lowary","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Alberta","place":["Edmonton, Canada"]},{"name":"Institute of Biological Chemistry, Academia Sinica","place":["Nangang, Taipei, Taiwan"]},{"name":"Institute of Biochemical Sciences, National Taiwan University","place":["Taipei, Taiwan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9454-5586","authenticated-orcid":false,"given":"Matthew S.","family":"Kimber","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph","place":["Guelph, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2025,10,30]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2021.755577"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwx034"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.carres.2007.12.012"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-genet-110410-132549"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2990"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.genet.42.110807.091427"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2003.03492.x"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.2002.tb00597.x"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2021.796045"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.182.19.5317-5324.2000"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.174.18.5941-5952.1992"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2007.01541.x"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.3.1537-1544.2004"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1094\/MPMI.1997.10.7.891"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1094\/MPMI-7-0099"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.23.6681-6689.2002"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.168.3.1392-1401.1986"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1080\/15592324.2015.1120396"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2001.02225.x"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M611669200"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M208824200"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwac079"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.9b00040"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.26528-0"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.3390\/biom11121795"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00374-003-0682-6"},{"key":"e_1_3_4_27_2","doi-asserted-by":"crossref","unstructured":"M. Hungria M. A. T. Vargas R. J. Campo L. M. O. Chueire D. S. de Andrade \u201cThe Brazilian experience with the soybean (Glycine max) and common bean (Phaseolus vulgaris) symbioses\u201d in Nitrogen Fixation: From Molecules to Crop Productivity F. O. Pedrosa M. Hungria G. Yates W. E. Newton Eds. (Springer 2000) pp. 515\u2013518.","DOI":"10.1007\/0-306-47615-0_292"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-13-735"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(97)00762-X"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.180.4.998-1001.1998"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)52212-2"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2002.03331.x"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1993.tb17607.x"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1006912121278"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.35.25069"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/0008-6215(95)00178-V"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/0008-6215(95)00300-2"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwz066"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0705709104"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-022-01488-1"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1106469"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2007.05.022"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkac387"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1107\/S1399004714001308"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1994.1303"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006067"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0730-x"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi990270y"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2020.06.016"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2008.08.121"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-00187-2"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M007399200"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2005.06.071"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-33988-1"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb950"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-019-0350-2"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.545384"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2011.10.039"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.76.061005.092322"},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.18388\/abp.2007_3227"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444911053157"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M610285200"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2024.1396268"},{"key":"e_1_3_4_64_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2003.03904.x"},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi3015554"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1997.1280"},{"key":"e_1_3_4_67_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-060713-035600"},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-022-01219-9"},{"key":"e_1_3_4_69_2","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-890-0:571"},{"key":"e_1_3_4_70_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10719-012-9451-9"},{"key":"e_1_3_4_71_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889807021206"},{"key":"e_1_3_4_72_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444902016657"},{"key":"e_1_3_4_73_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"e_1_3_4_74_2","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-381270-4.00019-6"},{"key":"e_1_3_4_75_2","doi-asserted-by":"crossref","unstructured":"T. J. B. Forrester M. S. Kimber WrtF fucosyltransferase \u2013 Apo. Protein Data Bank. https:\/\/www.rcsb.org\/structure\/9OMH. Deposited 14 May 2025.","DOI":"10.2210\/pdb9omh\/pdb"},{"key":"e_1_3_4_76_2","doi-asserted-by":"crossref","unstructured":"T. J. B. Forrester M. S. Kimber WrtF fucosyltransferase \u2013 GDP. Protein Data Bank. https:\/\/www.rcsb.org\/structure\/9OMI. Deposited 14 May 2025.","DOI":"10.2210\/pdb9omi\/pdb"},{"key":"e_1_3_4_77_2","doi-asserted-by":"crossref","unstructured":"T. J. B. Forrester M. S. Kimber WrtF fucosyltransferase \u2013 GDP-beta-l-fucose. Protein Data Bank. https:\/\/www.rcsb.org\/structure\/9OMJ. Deposited 14 May 2025.","DOI":"10.2210\/pdb9omj\/pdb"},{"key":"e_1_3_4_78_2","doi-asserted-by":"crossref","unstructured":"T. J. B. Forrester M. S. Kimber WrtF fucosyltransferase \u2013 trisaccharide acceptor. Protein Data Bank. https:\/\/www.rcsb.org\/structure\/9OMK. Deposited 14 May 2025.","DOI":"10.2210\/pdb9omk\/pdb"},{"key":"e_1_3_4_79_2","doi-asserted-by":"crossref","unstructured":"T. J. B. Forrester M. S. Kimber WrtF fucosyltransferase \u2013 acetylated trisaccharide acceptor. Protein Data Bank. https:\/\/www.rcsb.org\/structure\/9OML. Deposited 14 May 2025.","DOI":"10.2210\/pdb9oml\/pdb"},{"key":"e_1_3_4_80_2","doi-asserted-by":"crossref","unstructured":"T. J. B. Forrester M. S. Kimber WrtF fucosyltransferase \u2013 tetrasaccharide product. Protein Data Bank. https:\/\/www.rcsb.org\/structure\/9OMM. Deposited 14 May 2025.","DOI":"10.2210\/pdb9omm\/pdb"},{"key":"e_1_3_4_81_2","doi-asserted-by":"crossref","unstructured":"T. J. B. Forrester M. S. Kimber WrtF fucosyltransferase \u2013 GDP-2-deoxy-2-fluoro-beta-l-fucose. Protein Data Bank. https:\/\/www.rcsb.org\/structure\/9OMN. Deposited 14 May 2025.","DOI":"10.2210\/pdb9omn\/pdb"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2512460122","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T17:43:14Z","timestamp":1761846194000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2512460122"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,30]]},"references-count":81,"journal-issue":{"issue":"44","published-print":{"date-parts":[[2025,11,4]]}},"alternative-id":["10.1073\/pnas.2512460122"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2512460122","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,30]]},"assertion":[{"value":"2025-05-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-03","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-30","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2512460122"}}