{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,15]],"date-time":"2026-03-15T03:52:24Z","timestamp":1773546744766,"version":"3.50.1"},"reference-count":34,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2018,5,1]],"date-time":"2018-05-01T00:00:00Z","timestamp":1525132800000},"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":[[2018,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            GES-type \u03b2-lactamases are a group of enzymes that have evolved their hydrolytic activity against carbapenems. In this study, the role of residue 174 inside the \u03a9-loop of GES-1 and GES-5 was investigated. GES-1\n            <jats:sup>P174E<\/jats:sup>\n            and GES-5\n            <jats:sup>P174E<\/jats:sup>\n            mutants, selected by site saturation mutagenesis, were purified and kinetically characterized. In comparison with GES-1 and GES-5 wild-type enzymes, GES-1\n            <jats:sup>P174E<\/jats:sup>\n            and GES-5\n            <jats:sup>P174E<\/jats:sup>\n            mutants exhibited lower\n            <jats:italic>k<\/jats:italic>\n            <jats:sub>cat<\/jats:sub>\n            and\n            <jats:italic>k<\/jats:italic>\n            <jats:sub>cat<\/jats:sub>\n            \/\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            values for cephalosporins and penicillins. Concerning carbapenems, GES-1\n            <jats:sup>P174E<\/jats:sup>\n            shared higher\n            <jats:italic>k<\/jats:italic>\n            <jats:sub>cat<\/jats:sub>\n            values but lower\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            values than those calculated for GES-1. The GES-1\n            <jats:sup>P174E<\/jats:sup>\n            and GES-5\n            <jats:sup>P174E<\/jats:sup>\n            mutants showed high hydrolytic efficiency for imipenem, with\n            <jats:italic>k<\/jats:italic>\n            <jats:sub>cat<\/jats:sub>\n            \/\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            values 100- and 660-fold higher, respectively, than those of GES-1. Clavulanic acid and tazobactam are good inhibitors for both GES-1\n            <jats:sup>P174E<\/jats:sup>\n            and GES-5\n            <jats:sup>P174E<\/jats:sup>\n            . Molecular dynamic (MD) simulations carried out for GES-1, GES-5, GES-1\n            <jats:sup>P174E<\/jats:sup>\n            , and GES-5\n            <jats:sup>P174E<\/jats:sup>\n            complexed with imipenem and meropenem have shown that mutation at position 174 induces a drastic increase of enzyme flexibility, in particular in the \u03a9-loop. The circular dichroism (CD) spectroscopy spectra of the four enzymes indicate that the P174E substitution in GES-1 and GES-5 does not affect the secondary structural content of the enzymes.\n          <\/jats:p>","DOI":"10.1128\/aac.01851-17","type":"journal-article","created":{"date-parts":[[2018,3,1]],"date-time":"2018-03-01T13:05:53Z","timestamp":1519909553000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":12,"title":["P174E Substitution in GES-1 and GES-5 \u03b2-Lactamases Improves Catalytic Efficiency toward Carbapenems"],"prefix":"10.1128","volume":"62","author":[{"given":"Alessandra","family":"Piccirilli","sequence":"first","affiliation":[{"name":"Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Universit\u00e0 degli Studi dell'Aquila, L'Aquila, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paola Sandra","family":"Mercuri","sequence":"additional","affiliation":[{"name":"Laboratoire de Macromol\u00e9cules Biologiques, Centre d'Ing\u00e9nierie des Prot\u00e9ines, Universit\u00e9 de Li\u00e8ge, Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moreno","family":"Galleni","sequence":"additional","affiliation":[{"name":"Laboratoire de Macromol\u00e9cules Biologiques, Centre d'Ing\u00e9nierie des Prot\u00e9ines, Universit\u00e9 de Li\u00e8ge, Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Massimiliano","family":"Aschi","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Fisiche e Chimiche, Universit\u00e0 degli Studi dell'Aquila, L'Aquila, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andr\u00e9","family":"Matagne","sequence":"additional","affiliation":[{"name":"Laboratoire d'Enzymologie et Repliement des Prot\u00e9ines, Centre d'Ing\u00e9nierie des Prot\u00e9ines, Universit\u00e9 de Li\u00e8ge, Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gianfranco","family":"Amicosante","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Universit\u00e0 degli Studi dell'Aquila, L'Aquila, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariagrazia","family":"Perilli","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Universit\u00e0 degli Studi dell'Aquila, L'Aquila, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.3.622-632.2000"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01784-12"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00871-10"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1089\/mdr.2016.0152"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00773-10"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00072-09"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00360-16"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.49.1.474-475.2005"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2334-12-176"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2014.07.003"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444907036955"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja308197j"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi501052t"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0134(199701)27:1<47::AID-PROT6>3.0.CO;2-K"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.47.3.1062-1067.2003"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M116.749648"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01292-10"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1097(96)00288-1"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.47.12.3881-3889.2003"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(89)90358-2"},{"key":"e_1_3_2_22_2","volume-title":"Document M07-A9","author":"Clinical and Laboratory Standards Institute","year":"2012","unstructured":"Clinical and Laboratory Standards Institute. 2012. 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