{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T19:20:04Z","timestamp":1770664804156,"version":"3.49.0"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2006,12,1]],"date-time":"2006-12-01T00:00:00Z","timestamp":1164931200000},"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":[[2006,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Among the TEM-type extended-spectrum \u03b2-lactamases (ESBLs), an amino acid change at Ambler position 104 (Glu to Lys) results in increased resistance to ceftazidime and cefotaxime when found with other substitutions (e.g., Gly238Ser and Arg164Ser). To examine the role of Asp104 in SHV \u03b2-lactamases, site saturation mutagenesis was performed. Our goal was to investigate the properties of amino acid residues at this position that affect resistance to penicillins and oxyimino-cephalosporins. Unexpectedly, 58% of amino acid variants at position 104 in SHV expressed in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            DH10B resulted in \u03b2-lactamases with lowered resistance to ampicillin. In contrast, increased resistance to cefotaxime was demonstrated only for the Asp104Arg and Asp104Lys \u03b2-lactamases. When all 19 substitutions were introduced into the SHV-2 (Gly238Ser) ESBL, the most significant increases in cefotaxime and ceftazidime resistance were noted for both the doubly substituted Asp104Lys Gly238Ser and the doubly substituted Asp104Arg Gly238Ser \u03b2-lactamases. Correspondingly, the overall catalytic efficiency (\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            ) of hydrolysis for cefotaxime was increased from 0.60 \u00b1 0.07 \u03bcM\n            <jats:sup>\u22121<\/jats:sup>\n            s\n            <jats:sup>\u22121<\/jats:sup>\n            (mean \u00b1 standard deviation) for Gly238Ser to 1.70 \u00b1 0.01 \u03bcM\n            <jats:sup>\u22121<\/jats:sup>\n            s\n            <jats:sup>\u22121<\/jats:sup>\n            for the Asp104Lys and Gly238Ser \u03b2-lactamase (threefold increase). We also showed that (i)\n            <jats:italic>k<\/jats:italic>\n            <jats:sub>3<\/jats:sub>\n            was the rate-limiting step for the hydrolysis of cefotaxime by Asp104Lys, (ii) the\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            for cefotaxime of the doubly substituted Asp104Lys Gly238Ser variant approached that of the Gly238Ser \u03b2-lactamase as pH increased, and (iii) Lys at position 104 functions in an energetically additive manner with the Gly238Ser substitution to enhance catalysis of cephalothin. Based on this analysis, we propose that the amino acid at Ambler position 104 in SHV-1 \u03b2-lactamase plays a major role in substrate binding and recognition of oxyimino-cephalosporins and influences the interactions of Tyr105 with penicillins.\n          <\/jats:p>","DOI":"10.1128\/aac.00848-06","type":"journal-article","created":{"date-parts":[[2006,11,22]],"date-time":"2006-11-22T19:39:16Z","timestamp":1164224356000},"page":"4124-4131","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":33,"title":["Role of Asp104 in the SHV \u03b2-Lactamase"],"prefix":"10.1128","volume":"50","author":[{"given":"Christopher R.","family":"Bethel","sequence":"first","affiliation":[{"name":"Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrea M.","family":"Hujer","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kristine M.","family":"Hujer","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jodi M.","family":"Thomson","sequence":"additional","affiliation":[{"name":"Departments of Pharmacology"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark W.","family":"Ruszczycky","sequence":"additional","affiliation":[{"name":"Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vernon E.","family":"Anderson","sequence":"additional","affiliation":[{"name":"Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marianne","family":"Pusztai-Carey","sequence":"additional","affiliation":[{"name":"Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Magdalena","family":"Taracila","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marion S.","family":"Helfand","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio"},{"name":"Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, Ohio"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert A.","family":"Bonomo","sequence":"additional","affiliation":[{"name":"Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio"},{"name":"Departments of Pharmacology"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj2760269"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(02)00599-5"},{"key":"e_1_3_2_4_2","unstructured":"Bell R. 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