{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T22:17:36Z","timestamp":1772662656474,"version":"3.50.1"},"reference-count":32,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[1999,8,1]],"date-time":"1999-08-01T00:00:00Z","timestamp":933465600000},"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":[[1999,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Extended-spectrum TEM \u03b2-lactamases (ESBLs) do not usually confer resistance to \u03b2-lactamase inhibitors such as clavulanate or tazobactam. To investigate the compatibility of the two phenotypes we used site-directed mutagenesis of the\n            <jats:italic>bla<\/jats:italic>\n            <jats:sub>TEM-1<\/jats:sub>\n            gene to introduce into the TEM-1 \u03b2-lactamase amino acid substitutions that confer the ESBL phenotype: TEM-12 (Arg164\u2192Ser), TEM-26 (Arg164\u2192Ser plus Glu104\u2192Lys), TEM-19 (Gly238\u2192Ser), and TEM-15 (Gly238\u2192Ser plus Glu104\u2192Lys). These were combined with three sets of substitutions that confer inhibitor resistance: TEM-31 (Arg244\u2192Cys), TEM-33 (Met69\u2192Leu), and TEM-35 (Met69\u2192Leu and Asn276\u2192Asp). Introduction of the Arg244\u2192Cys substitution gave rise to inhibitor-resistant hybrid enzymes that either lost ESBL activity (TEM-12, TEM-15, and TEM-19) or had reduced activity (TEM-26) against ceftazidime. In contrast, the introduction of Met69\u2192Leu or Met69\u2192Leu plus Asn276\u2192Asp substitutions did not significantly affect the abilities of the enzymes to confer resistance to ceftazidime, although increased susceptibility to cefotaxime was observed with\n            <jats:italic>Escherichia coli<\/jats:italic>\n            strains that expressed the TEM-19 and TEM-26 \u03b2-lactamases. With the exception of the TEM-12 \u03b2-lactamase, introduction of the Met69\u2192Leu substitution did not give rise to enzymes with increased resistance to clavulanate compared to that of the TEM-1 \u03b2-lactamase. However, introduction of the double substitution Met69\u2192Leu plus Asn276\u2192Asp in the ESBLs did give rise to low-level (TEM-19, TEM-15, and TEM-26) or moderate-level (TEM-12) clavulanate resistance. None of the hybrid enzymes were as resistant to clavulanate as the corresponding inhibitor-resistant TEM \u03b2-lactamase mutant, suggesting that active-site configuration in the ESBLs limits the degree of clavulanate resistance conferred.\n          <\/jats:p>","DOI":"10.1128\/aac.43.8.1881","type":"journal-article","created":{"date-parts":[[2018,10,9]],"date-time":"2018-10-09T00:35:20Z","timestamp":1539045320000},"page":"1881-1887","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":24,"title":["Construction and Characterization of Mutants of the TEM-1 \u03b2-Lactamase Containing Amino Acid Substitutions Associated with both Extended-Spectrum Resistance and Resistance to \u03b2-Lactamase Inhibitors"],"prefix":"10.1128","volume":"43","author":[{"given":"Paul D.","family":"Stapleton","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Department of Microbiology, The Guy\u2019s, King\u2019s &amp; St. Thomas\u2019 School of Medicine, St. Thomas\u2019 Campus, London SE1 7EH, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin P.","family":"Shannon","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Microbiology, The Guy\u2019s, King\u2019s &amp; St. Thomas\u2019 School of Medicine, St. Thomas\u2019 Campus, London SE1 7EH, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gary L.","family":"French","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Microbiology, The Guy\u2019s, King\u2019s &amp; St. Thomas\u2019 School of Medicine, St. Thomas\u2019 Campus, London SE1 7EH, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"75","article-title":"Nucleotide sequences of the genes coding for the TEM-like \u03b2-lactamases IRT-1 and IRT-2 (formerly called TRI-1 and TRI-2).","volume":"120","author":"Belaaouaj A.","year":"1994","unstructured":"Belaaouaj A. Lapoumeroulie C. Canica M. M. Vedel G. N\u00e9vot P. Krishnamoorthy R. Paul G. Nucleotide sequences of the genes coding for the TEM-like \u03b2-lactamases IRT-1 and IRT-2 (formerly called TRI-1 and TRI-2). FEMS Microbiol. Lett. 120 1994 75 80","journal-title":"FEMS Microbiol. Lett."},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.9.1940"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.11.2547"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/38.2.183"},{"key":"e_1_3_2_6_2","first-page":"111","article-title":"Characterization and amino acid sequence of IRT-4, a novel TEM-type enzyme with a decreased susceptibility to \u03b2-lactamase inhibitors.","volume":"120","author":"Brun T.","year":"1994","unstructured":"Brun T. P\u00e9duzzi J. Cani\u00e7a M. M. Paul G. N\u00e9vot P. Barth\u00e9l\u00e9my M. Labia R. Characterization and amino acid sequence of IRT-4, a novel TEM-type enzyme with a decreased susceptibility to \u03b2-lactamase inhibitors. FEMS Microbiol. Lett. 120 1994 111 118","journal-title":"FEMS Microbiol. Lett."},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.6.1211"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.6.1323"},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"668","DOI":"10.1093\/jac\/39.5.668","article-title":"Are TEM \u03b2-lactamases encoded by pBR322 and Bluescript plasmids enzymatically indistinguishable?","volume":"39","author":"Chaibi E. B.","year":"1997","unstructured":"Chaibi E. B. P\u00e9duzzi J. Barth\u00e9l\u00e9my M. Labia R. Are TEM \u03b2-lactamases encoded by pBR322 and Bluescript plasmids enzymatically indistinguishable? J. Antimicrob. Chemother. 39 1997 668 669","journal-title":"J. Antimicrob. Chemother."},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-4838(97)00127-1"},{"key":"e_1_3_2_11_2","doi-asserted-by":"crossref","first-page":"20600","DOI":"10.1016\/S0021-9258(19)36729-8","article-title":"Site-directed mutagenesis at the active site of Escherichia coli TEM-1 \u03b2-lactamase. Suicide inhibitor-resistant mutants reveal the role of arginine 244 and methionine 69 in catalysis.","volume":"267","author":"Delaire M.","year":"1992","unstructured":"Delaire M. Labia R. Samama J. P. Masson J. M. Site-directed mutagenesis at the active site of Escherichia coli TEM-1 \u03b2-lactamase. Suicide inhibitor-resistant mutants reveal the role of arginine 244 and methionine 69 in catalysis. J. Biol. Chem. 267 1992 20600 20606","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.40.10.2434"},{"key":"e_1_3_2_13_2","doi-asserted-by":"crossref","first-page":"1281","DOI":"10.1128\/AAC.42.5.1281","article-title":"Properties of mutant SHV-5 \u03b2-lactamases constructed by substitution of isoleucine or valine for methionine at position 69.","volume":"42","author":"Giakkoup P.","year":"1998","unstructured":"Giakkoup P. Miriagou V. Gazouli M. Tzelepi E. Legakis N. J. Tzouvelekis L. S. Properties of mutant SHV-5 \u03b2-lactamases constructed by substitution of isoleucine or valine for methionine at position 69. Antimicrob. Agents Chemother. 42 1998 1281 1283","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.2.427"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja00064a003"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.38.5.1134"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.12.2593"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(87)54085-X"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.8.4.557"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj3300581"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.8153625"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.31.18240"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.6.1322"},{"key":"e_1_3_2_24_2","doi-asserted-by":"crossref","first-page":"3179","DOI":"10.1021\/bi00227a004","article-title":"Substitution of lysine at position 104 or 240 of TEM-1pTZ18R \u03b2-lactamase enhances the effect of serine-164 substitution on hydrolysis or affinity for cephalosporins and the monobactam aztreonam.","volume":"30","author":"Sowek J. A.","year":"1991","unstructured":"Sowek J. A. Singer S. B. Ohringer S. Malley M. F. Dougherty T. J. Gougoutas J. Z. Bush K. Substitution of lysine at position 104 or 240 of TEM-1pTZ18R \u03b2-lactamase enhances the effect of serine-164 substitution on hydrolysis or affinity for cephalosporins and the monobactam aztreonam. Biochemistry 30 1991 3179 3188","journal-title":"Biochemistry"},{"key":"e_1_3_2_25_2","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1126\/science.8153626","article-title":"Resistance to antibiotics mediated by target alterations.","volume":"265","author":"Spratt B. G.","year":"1994","unstructured":"Spratt B. G. Resistance to antibiotics mediated by target alterations. Science 265 1994 388 393","journal-title":"Science"},{"key":"e_1_3_2_26_2","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1093\/jac\/36.3.483","article-title":"The ability of \u03b2-lactam antibiotics to select mutants with derepressed \u03b2-lactamase synthesis from Citrobacter freundii.","volume":"36","author":"Stapleton P.","year":"1995","unstructured":"Stapleton P. Shannon K. Phillips I. The ability of \u03b2-lactam antibiotics to select mutants with derepressed \u03b2-lactamase synthesis from Citrobacter freundii. J. Antimicrob. Chemother. 36 1995 483 496","journal-title":"J. Antimicrob. Chemother."},{"key":"e_1_3_2_27_2","doi-asserted-by":"crossref","first-page":"2479","DOI":"10.1128\/AAC.39.11.2478","article-title":"Incidence and mechanisms of resistance to the combination of amoxicillin and clavulanic acid in Escherichia coli.","volume":"39","author":"Stapleton P.","year":"1995","unstructured":"Stapleton P. Wu P.-J. King A. Shannon K. French G. Phillips I. 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Clinical isolates of Escherichia coli producing TRI \u03b2-lactamases: novel TEM-enzymes conferring resistance to \u03b2-lactamase inhibitors. J. Antimicrob. Chemother. 30 1992 449 462","journal-title":"J. Antimicrob. 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