{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T01:27:28Z","timestamp":1789954048414,"version":"4.0.1"},"reference-count":44,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2013,4,1]],"date-time":"2013-04-01T00:00:00Z","timestamp":1364774400000},"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":[[2013,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Class C cephalosporinases are a growing threat, and clinical inhibitors of these enzymes are currently unavailable. Previous studies have explored the role of Asn152 in the\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Escherichia coli<\/jats:named-content>\n            AmpC and P99 enzymes and have suggested that interactions between C-6\u2032 or C-7\u2032 substituents on penicillins or cephalosporins and Asn152 are important in determining substrate specificity and enzymatic stability. We sought to characterize the role of Asn152 in the clinically important CMY-2 cephalosporinase with substrates and inhibitors. Mutagenesis of CMY-2 at position 152 yields functional mutants (N152G, -S, and -T) that exhibit improved penicillinase activity and retain cephamycinase activity. We also tested whether the position 152 substitutions would affect the inactivation kinetics of tazobactam, a class A \u03b2-lactamase inhibitor with\n            <jats:italic>in vitro<\/jats:italic>\n            activity against CMY-2. Using standard assays, we showed that the N152G, -S, and -T variants possessed increased catalytic activity against cefoxitin compared to the wild type. The 50% inhibitory concentration (IC\n            <jats:sub>50<\/jats:sub>\n            ) for tazobactam improved dramatically, with an 18-fold reduction for the N152S mutant due to higher rates of enzyme inactivation. Modeling studies have shown active-site expansion due to interactions between Y150 and S152 in the apoenzyme and the Michaelis-Menten complex with tazobactam. Substitutions at N152 might become clinically important as new class C \u03b2-lactamase inhibitors are developed.\n          <\/jats:p>","DOI":"10.1128\/aac.01334-12","type":"journal-article","created":{"date-parts":[[2013,1,15]],"date-time":"2013-01-15T04:53:42Z","timestamp":1358225622000},"page":"1596-1602","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":7,"title":["N152G, -S, and -T Substitutions in CMY-2 \u03b2-Lactamase Increase Catalytic Efficiency for Cefoxitin and Inactivation Rates for Tazobactam"],"prefix":"10.1128","volume":"57","author":[{"given":"Marion J.","family":"Skalweit","sequence":"first","affiliation":[{"name":"Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"},{"name":"Departments of Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"},{"name":"Biochemistry, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei","family":"Li","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin C.","family":"Conklin","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magdalena A.","family":"Taracila","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"},{"name":"Departments of Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca A.","family":"Hutton","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00036-08"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00810-07"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.43.2.424"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.36.9.2046"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.50.5.1633-1641.2006"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.2217\/fmb.11.128"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00050-10"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijantimicag.2008.04.019"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00133-09"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi9015549"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00219-09"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/prot.23150"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.05630-11"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.48.7.2652-2658.2004"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00023a023"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.073092407"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1089\/10766290260469598"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M207271200"},{"key":"e_1_3_3_20_2","volume-title":"CLSI document M07-A9","author":"Clinical and Laboratory Standards Institute, CLSI","year":"2012","unstructured":"Clinical and Laboratory Standards Institute, CLSI. 2012. 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Exploring the role of Asn 152 in altering the resistance spectrum of the class C \u03b2-lactamase CMY-2, poster 482. Abstr. 243rd American Chemical Society National Meeting & Exposition. 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