{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T09:35:16Z","timestamp":1740130516013,"version":"3.37.3"},"reference-count":39,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2017,7,21]],"date-time":"2017-07-21T00:00:00Z","timestamp":1500595200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["CIHR MOP\u2010114889"],"award-info":[{"award-number":["CIHR MOP\u2010114889"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2017,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The aminoglycoside modifying enzyme (AME) ANT(2\u2033)\u2010Ia is a significant target for next generation antibiotic development. Structural studies of a related aminoglycoside\u2010modifying enzyme, ANT(3\u2033)(9), revealed this enzyme contains dynamic, disordered, and well\u2010defined segments that modulate thermodynamically before and after antibiotic binding. Characterizing these structural dynamics is critical for <jats:italic>in situ<\/jats:italic> screening, design, and development of contemporary antibiotics that can be implemented in a clinical setting to treat potentially lethal, antibiotic resistant, human infections. Here, the first NMR structural ensembles of ANT(2\u2033)\u2010Ia are presented, and suggest that ATP\u2010aminoglycoside binding repositions the nucleotidyltransferase (NT) and C\u2010terminal domains for catalysis to efficiently occur. Residues involved in ligand recognition were assessed by site\u2010directed mutagenesis. <jats:italic>In vitro<\/jats:italic> activity assays indicate a critical role for I129 toward aminoglycoside modification in addition to known catalytic D44, D46, and D48 residues. These observations support previous claims that ANT aminoglycoside sub\u2010class promiscuity is not solely due to binding cleft size, or inherent partial disorder, but can be controlled by ligand modulation on distinct dynamic and thermodynamic properties of ANTs under cellular conditions. Hydrophobic interactions in the substrate binding cleft, as well as solution dynamics in the C\u2010terminal tail of ANT(2\u2033)\u2010Ia, advocate toward design of kanamycin\u2010derived cationic lipid aminoglycoside analogs, some of which have already shown antimicrobial activity <jats:italic>in vivo<\/jats:italic> against kanamycin and gentamicin\u2010resistant <jats:italic>P. aeruginosa<\/jats:italic>. This data will drive additional <jats:italic>in silico<\/jats:italic>, next generation antibiotic development for future human use to combat increasingly prevalent antimicrobial resistance.<\/jats:p>","DOI":"10.1002\/pro.3224","type":"journal-article","created":{"date-parts":[[2017,7,22]],"date-time":"2017-07-22T03:16:46Z","timestamp":1500693406000},"page":"1852-1863","source":"Crossref","is-referenced-by-count":2,"title":["Effect of solvent and protein dynamics in ligand recognition and inhibition of aminoglycoside adenyltransferase 2\u2033\u2010Ia"],"prefix":"10.1002","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6620-6560","authenticated-orcid":false,"given":"Valjean R.","family":"Bacot\u2010Davis","sequence":"first","affiliation":[{"name":"McGill University, Biochemistry 3649 Promenade Sir William Osler Room 470 Montreal QC H3A 0G4 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angelia V.","family":"Bassenden","sequence":"additional","affiliation":[{"name":"McGill University, Biochemistry 3649 Promenade Sir William Osler Room 470 Montreal QC H3A 0G4 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tara","family":"Sprules","sequence":"additional","affiliation":[{"name":"McGill University, Biochemistry 3649 Promenade Sir William Osler Room 470 Montreal QC H3A 0G4 Canada"},{"name":"Quebec\/Eastern Canada NMR Centre, Pulp &amp; Paper Research Centre 3420 University St. Room 023 Montreal QC H3A 2A7 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Albert M.","family":"Berghuis","sequence":"additional","affiliation":[{"name":"McGill University, Biochemistry 3649 Promenade Sir William Osler Room 470 Montreal QC H3A 0G4 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,7,21]]},"reference":[{"volume-title":"Textbook of adult emergency 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