{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:52:49Z","timestamp":1786978369242,"version":"build-2736575974"},"reference-count":29,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2020,1,29]],"date-time":"2020-01-29T00:00:00Z","timestamp":1580256000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["HHSN272201200026C"],"award-info":[{"award-number":["HHSN272201200026C"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["HHSN272201700060C"],"award-info":[{"award-number":["HHSN272201700060C"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["HHSN272200700058C"],"award-info":[{"award-number":["HHSN272200700058C"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2020,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Aminoglycosides were one of the first classes of broad\u2010spectrum antibacterial drugs clinically used to effectively combat infections. The rise of resistance to these drugs, mediated by enzymatic modification, has since compromised their utility as a treatment option, prompting intensive research into the molecular function of resistance enzymes. Here, we report the crystal structure of aminoglycoside nucleotidyltransferase ANT(4\u2032)\u2010IIb in apo and tobramycin\u2010bound forms at a resolution of 1.6 and 2.15\u2009\u00c5, respectively. ANT(4\u2032)\u2010IIb was discovered in the opportunistic pathogen\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                    <\/jats:styled-content>\n                    and conferred resistance to amikacin and tobramycin. Analysis of the ANT(4\u2032)\u2010IIb structures revealed a two\u2010domain organization featuring a mixed \u03b2\u2010sheet and an \u03b1\u2010helical bundle. ANT(4\u2032)\u2010IIb monomers form a dimer required for its enzymatic activity, as coordination of the aminoglycoside substrate relies on residues contributed by both monomers. Despite harbouring appreciable primary sequence diversity compared to previously characterized homologues, the ANT(4\u2032)\u2010IIb structure demonstrates a surprising level of structural conservation highlighting the high plasticity of this general protein fold. Site\u2010directed mutagenesis of active site residues and kinetic analysis provides support for a catalytic mechanism similar to those of other nucleotidyltransferases. Using the molecular insights provided into this ANT(4\u2032)\u2010IIb\u2010represented enzymatic group, we provide a hypothesis for the potential evolutionary origin of these aminoglycoside resistance determinants.\n                  <\/jats:p>","DOI":"10.1002\/pro.3815","type":"journal-article","created":{"date-parts":[[2019,12,31]],"date-time":"2019-12-31T08:36:00Z","timestamp":1577781360000},"page":"758-767","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Structural characterization of aminoglycoside 4\u2032\u2010\n                    <i>O<\/i>\n                    \u2010adenylyltransferase ANT(4\u2032)\u2010IIb from\n                    <i>Pseudomonas aeruginosa<\/i>"],"prefix":"10.1002","volume":"29","author":[{"given":"Cameron","family":"Semper","sequence":"first","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Disease University of Calgary  Calgary Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Stogios","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Applied Chemistry University of Toronto  Toronto Ontario Canada"},{"name":"Center for Structural Genomics of Infectious Diseases (CSGID)"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Djalal","family":"Meziane\u2010Cherif","sequence":"additional","affiliation":[{"name":"Institut Pasteur Unit\u00e9 des Agents Anitbact\u00e9riens  Paris Cedex France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Evdokimova","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Applied Chemistry University of Toronto  Toronto Ontario Canada"},{"name":"Center for Structural Genomics of Infectious Diseases (CSGID)"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrice","family":"Courvalin","sequence":"additional","affiliation":[{"name":"Institut Pasteur Unit\u00e9 des Agents Anitbact\u00e9riens  Paris Cedex France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5256-9237","authenticated-orcid":false,"given":"Alexei","family":"Savchenko","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Disease University of Calgary  Calgary Alberta Canada"},{"name":"Department of Chemical Engineering and Applied Chemistry University of Toronto  Toronto Ontario Canada"},{"name":"Center for Structural Genomics of Infectious Diseases 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