{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T19:30:55Z","timestamp":1776022255007,"version":"3.50.1"},"reference-count":19,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2013,8,1]],"date-time":"2013-08-01T00:00:00Z","timestamp":1375315200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2013,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The aminoglycoside 2\u2033-phosphotransferases APH(2\u2033)-IIa and APH(2\u2033)-IVa can utilize ATP and GTP as cosubstrates, since both enzymes possess overlapping but discrete structural templates for ATP and GTP binding. APH(2\u2033)-IIIa uses GTP exclusively, because its ATP-binding template is blocked by a bulky tyrosine \u201cgatekeeper\u201d residue. Replacement of the \u201cgatekeeper\u201d residues M85 and F95 in APH(2\u2033)-IIa and APH(2\u2033)-IVa, respectively, by tyrosine does not significantly change the antibiotic susceptibility profiles produced by the enzymes. In APH(2\u2033)-IIa, M85Y substitution results in an \u223c10-fold decrease in the\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            value of GTP and an \u223c320-fold increase in the\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            value of ATP. In APH(2\u2033)-IVa, F95Y substitution results in a modest decrease in the\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            values of both GTP and ATP. Structural analysis indicates that in the APH(2\u2033)-IIa M85Y mutant, tyrosine blocks access of ATP to the correct position in the binding site, while the larger nucleoside triphosphate (NTP)-binding pocket of the APH(2\u2033)-IVa F95Y mutant allows the tyrosine to move away, thus giving access to the ATP-binding template.\n          <\/jats:p>","DOI":"10.1128\/aac.00381-13","type":"journal-article","created":{"date-parts":[[2013,5,29]],"date-time":"2013-05-29T04:58:59Z","timestamp":1369803539000},"page":"3763-3766","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":5,"title":["Bulky \u201cGatekeeper\u201d Residue Changes the Cosubstrate Specificity of Aminoglycoside 2\u2033-Phosphotransferase IIa"],"prefix":"10.1128","volume":"57","author":[{"given":"Monolekha","family":"Bhattacharya","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marta","family":"Toth","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Clyde A.","family":"Smith","sequence":"additional","affiliation":[{"name":"Stanford Synchrotron Radiation Lightsource, Stanford University, Menlo Park, California, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sergei B.","family":"Vakulenko","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.16.3.430-450.2003"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.2741\/culebras"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.70.8.2276"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.167.2.631-638.1986"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.3.511"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.10.2876-2879.2000"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.5.1229"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M808148200"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.416453"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.186"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01020-07"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.437"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.349670"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.341206"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01444-09"},{"key":"e_1_3_2_17_2","volume-title":"Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically: approved standard","author":"CLSI","year":"2009","unstructured":"CLSI. 2009. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically: approved standard, 8th ed. Clinical and Laboratory Standards Institute, Wayne, PA.","edition":"8"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkg563"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi6024512"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00149-09"}],"container-title":["Antimicrobial Agents and Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.00381-13","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.00381-13","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,21]],"date-time":"2022-02-21T22:32:39Z","timestamp":1645482759000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AAC.00381-13"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,8]]},"references-count":19,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2013,8]]}},"alternative-id":["10.1128\/AAC.00381-13"],"URL":"https:\/\/doi.org\/10.1128\/aac.00381-13","relation":{},"ISSN":["0066-4804","1098-6596"],"issn-type":[{"value":"0066-4804","type":"print"},{"value":"1098-6596","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,8]]},"assertion":[{"value":"2013-02-21","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2013-05-19","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2013-07-15","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}