{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T17:19:24Z","timestamp":1789665564296,"version":"4.0.1"},"reference-count":25,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2019,2,6]],"date-time":"2019-02-06T00:00:00Z","timestamp":1549411200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["1SC3GM098173"],"award-info":[{"award-number":["1SC3GM098173"]}],"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":[[2019,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:italic>Pseudomonas aeruginosa<\/jats:italic> is an opportunistic multi\u2010drug resistant pathogen implicated as a causative agent in nosocomial and community acquired bacterial infections. The gene encoding prolyl\u2010tRNA synthetase (ProRS) from <jats:italic>P. aeruginosa<\/jats:italic> was overexpressed in <jats:italic>Escherichia coli<\/jats:italic> and the resulting protein was characterized. ProRS was kinetically evaluated and the <jats:italic>K<\/jats:italic><jats:sub>M<\/jats:sub> values for interactions with ATP, proline, and tRNA were 154, 122, and 5.5 \u03bcM, respectively. The turn\u2010over numbers, <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub><jats:sup>obs<\/jats:sup>, for interactions with these substrates were calculated to be 5.5, 6.3, and 0.2 s<jats:sup>\u22121<\/jats:sup>, respectively. The crystal structure of the \u03b1<jats:sub>2<\/jats:sub> form of <jats:italic>P. aeruginosa<\/jats:italic> ProRS was solved to 2.60\u2009\u00c5 resolution. The amino acid sequence and X\u2010ray crystal structure of <jats:italic>P. aeruginosa<\/jats:italic> ProRS was analyzed and compared with homologs in which the crystal structures have been solved. The amino acids that interact with ATP and proline are well conserved in the active site region and overlay of the crystal structure with ProRS homologs conforms to a similar overall three\u2010dimensional structure. ProRS was developed into a screening platform using scintillation proximity assay (SPA) technology and used to screen 890 chemical compounds, resulting in the identification of two inhibitory compounds, BT06A02 and BT07H05. This work confirms the utility of a screening system based on the functionality of ProRS from <jats:italic>P. aeruginosa<\/jats:italic>.<\/jats:p>","DOI":"10.1002\/pro.3579","type":"journal-article","created":{"date-parts":[[2019,1,22]],"date-time":"2019-01-22T05:01:26Z","timestamp":1548133286000},"page":"727-737","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Characterization and structure determination of prolyl\u2010tRNA synthetase from <i>Pseudomonas aeruginosa<\/i> and development as a screening platform"],"prefix":"10.1002","volume":"28","author":[{"given":"Noah","family":"Pena","sequence":"first","affiliation":[{"name":"The University of Texas \u2013 RGV  Edinburg Texas 78541"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M.","family":"Dranow","sequence":"additional","affiliation":[{"name":"Seattle Structural Genomics Center for Infectious Disease  Seattle WA 98109"},{"name":"Beryllium Discovery Corp  Bainbridge Island WA 98110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanmei","family":"Hu","sequence":"additional","affiliation":[{"name":"The University of Texas \u2013 RGV  Edinburg Texas 78541"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaritza","family":"Escamilla","sequence":"additional","affiliation":[{"name":"The University of Texas \u2013 RGV  Edinburg Texas 78541"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James M.","family":"Bullard","sequence":"additional","affiliation":[{"name":"The University of Texas \u2013 RGV  Edinburg Texas 78541"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2019,2,6]]},"reference":[{"key":"e_1_2_7_2_1","first-page":"1","volume-title":"Antibiotic Resistance: federal agencies need to better focus efforts to address risk to humans from antibiotic use in animals. 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