Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1996 Oct;40(10):2434–2436. doi: 10.1128/aac.40.10.2434

Implication of Ile-69 and Thr-182 residues in kinetic characteristics of IRT-3 (TEM-32) beta-lactamase.

S Farzaneh 1, E B Chaibi 1, J Peduzzi 1, M Barthelemy 1, R Labia 1, J Blazquez 1, F Baquero 1
PMCID: PMC163551  PMID: 8891161

Abstract

The substitution of a methionine for an isoleucine at position 69 (Met69Ile), which causes inhibitor resistance to TEM-type beta-lactamases (IRT-3 and IRT-I69), altered the positions of the Asn-170 and Glu-166 side chains as well as the position of the catalytic water molecule. A novel hydrogen bond between the hydroxyl of Thr-182 and the carbonyl of Glu-64 was expected to be responsible for the increase in the catalytic activity of the IST-T182 and IRT-3 enzymes compared with those of TEM-1 and IRT-169, respectively.

Full Text

The Full Text of this article is available as a PDF (164.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ambler R. P., Coulson A. F., Frère J. M., Ghuysen J. M., Joris B., Forsman M., Levesque R. C., Tiraby G., Waley S. G. A standard numbering scheme for the class A beta-lactamases. Biochem J. 1991 May 15;276(Pt 1):269–270. doi: 10.1042/bj2760269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Belaaouaj A., Lapoumeroulie C., Caniça M. M., Vedel G., Névot P., Krishnamoorthy R., Paul G. Nucleotide sequences of the genes coding for the TEM-like beta-lactamases IRT-1 and IRT-2 (formerly called TRI-1 and TRI-2). FEMS Microbiol Lett. 1994 Jul 1;120(1-2):75–80. doi: 10.1111/j.1574-6968.1994.tb07010.x. [DOI] [PubMed] [Google Scholar]
  3. Bonomo R. A., Currie-McCumber C., Shlaes D. M. OHIO-1 beta-lactamase resistant to mechanism-based inactivators. FEMS Microbiol Lett. 1992 Apr 1;71(1):79–82. doi: 10.1016/0378-1097(92)90545-y. [DOI] [PubMed] [Google Scholar]
  4. Bonomo R. A., Dawes C. G., Knox J. R., Shlaes D. M. Complementary roles of mutations at positions 69 and 242 in a class A beta-lactamase. Biochim Biophys Acta. 1995 Feb 22;1247(1):113–120. doi: 10.1016/0167-4838(94)00187-l. [DOI] [PubMed] [Google Scholar]
  5. Bonomo R. A., Dawes C. G., Knox J. R., Shlaes D. M. beta-Lactamase mutations far from the active site influence inhibitor binding. Biochim Biophys Acta. 1995 Feb 22;1247(1):121–125. doi: 10.1016/0167-4838(94)00188-m. [DOI] [PubMed] [Google Scholar]
  6. Brun T., Péduzzi J., Caniça M. M., Paul G., Névot P., Barthélémy M., Labia R. Characterization and amino acid sequence of IRT-4, a novel TEM-type enzyme with a decreased susceptibility to beta-lactamase inhibitors. FEMS Microbiol Lett. 1994 Jul 1;120(1-2):111–117. doi: 10.1111/j.1574-6968.1994.tb07016.x. [DOI] [PubMed] [Google Scholar]
  7. Delaire M., Labia R., Samama J. P., Masson J. M. Site-directed mutagenesis at the active site of Escherichia coli TEM-1 beta-lactamase. Suicide inhibitor-resistant mutants reveal the role of arginine 244 and methionine 69 in catalysis. J Biol Chem. 1992 Oct 15;267(29):20600–20606. [PubMed] [Google Scholar]
  8. Henquell C., Chanal C., Sirot D., Labia R., Sirot J. Molecular characterization of nine different types of mutants among 107 inhibitor-resistant TEM beta-lactamases from clinical isolates of Escherichia coli. Antimicrob Agents Chemother. 1995 Feb;39(2):427–430. doi: 10.1128/aac.39.2.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jelsch C., Mourey L., Masson J. M., Samama J. P. Crystal structure of Escherichia coli TEM1 beta-lactamase at 1.8 A resolution. Proteins. 1993 Aug;16(4):364–383. doi: 10.1002/prot.340160406. [DOI] [PubMed] [Google Scholar]
  10. Knox J. R. Extended-spectrum and inhibitor-resistant TEM-type beta-lactamases: mutations, specificity, and three-dimensional structure. Antimicrob Agents Chemother. 1995 Dec;39(12):2593–2601. doi: 10.1128/aac.39.12.2593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Labia R., Andrillon J., Le Goffic F. Computerized microacidimetric determination of beta lactamase Michaelis-Menten constants. FEBS Lett. 1973 Jun 15;33(1):42–44. doi: 10.1016/0014-5793(73)80154-1. [DOI] [PubMed] [Google Scholar]
  12. Oliphant A. R., Struhl K. An efficient method for generating proteins with altered enzymatic properties: application to beta-lactamase. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9094–9098. doi: 10.1073/pnas.86.23.9094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Péduzzi J., Reynaud A., Baron P., Barthélémy M., Labia R. Chromosomally encoded cephalosporin-hydrolyzing beta-lactamase of Proteus vulgaris RO104 belongs to Ambler's class A. Biochim Biophys Acta. 1994 Jul 20;1207(1):31–39. doi: 10.1016/0167-4838(94)90048-5. [DOI] [PubMed] [Google Scholar]
  14. Sirot D., Chanal C., Henquell C., Labia R., Sirot J., Cluzel R. Clinical isolates of Escherichia coli producing multiple TEM mutants resistant to beta-lactamase inhibitors. J Antimicrob Chemother. 1994 Jun;33(6):1117–1126. doi: 10.1093/jac/33.6.1117. [DOI] [PubMed] [Google Scholar]
  15. Stapleton P., Wu P. J., King A., Shannon K., French G., Phillips I. Incidence and mechanisms of resistance to the combination of amoxicillin and clavulanic acid in Escherichia coli. Antimicrob Agents Chemother. 1995 Nov;39(11):2478–2483. doi: 10.1128/aac.39.11.2478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sutcliffe J. G. Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3737–3741. doi: 10.1073/pnas.75.8.3737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Thomson C. J., Amyes S. G. Selection of variants of the TEM-1 beta-lactamase, encoded by a plasmid of clinical origin, with increased resistance to beta-lactamase inhibitors. J Antimicrob Chemother. 1993 May;31(5):655–664. doi: 10.1093/jac/31.5.655. [DOI] [PubMed] [Google Scholar]
  18. Thomson C. J., Amyes S. G. TRC-1: emergence of a clavulanic acid-resistant TEM beta-lactamase in a clinical strain. FEMS Microbiol Lett. 1992 Mar 1;70(2):113–117. doi: 10.1016/0378-1097(92)90669-f. [DOI] [PubMed] [Google Scholar]
  19. Vedel G., Belaaouaj A., Gilly L., Labia R., Philippon A., Névot P., Paul G. Clinical isolates of Escherichia coli producing TRI beta-lactamases: novel TEM-enzymes conferring resistance to beta-lactamase inhibitors. J Antimicrob Chemother. 1992 Oct;30(4):449–462. doi: 10.1093/jac/30.4.449. [DOI] [PubMed] [Google Scholar]
  20. Zhou X. Y., Bordon F., Sirot D., Kitzis M. D., Gutmann L. Emergence of clinical isolates of Escherichia coli producing TEM-1 derivatives or an OXA-1 beta-lactamase conferring resistance to beta-lactamase inhibitors. Antimicrob Agents Chemother. 1994 May;38(5):1085–1089. doi: 10.1128/aac.38.5.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES