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. 1994 Nov 1;303(Pt 3):825–830. doi: 10.1042/bj3030825

Kinetic and physical studies of beta-lactamase inhibition by a novel penem, BRL 42715.

T H Farmer 1, J W Page 1, D J Payne 1, D J Knowles 1
PMCID: PMC1137621  PMID: 7980451

Abstract

The interactions of Staphylococcus aureus, Bacillus cereus I, TEM, Klebsiella pneumoniae K1 and Enterobacter cloacae P99 beta-lactamases with the novel penem inhibitor BRL 42715 were investigated kinetically and, in some cases, by electrospray mass spectrometry (e.s.m.s.). All the beta-lactamases were rapidly inactivated by BRL 42715, with second-order rate constants ranging from 0.17 to 6.4 microM-1.s-1. The initial stoichiometry of beta-lactamase inhibition was essentially 1:1, with the exception of the K1 enzyme. In this instance about 20 molecules of BRL 42715 were hydrolysed before the enzyme was completely inhibited. Inhibited beta-lactamases did not readily regain activity in the absence of BRL 42715, the half-lives for regeneration of free enzyme ranging from 5 min for the K1 beta-lactamase to over 2 days for the staphylococcal enzyme. Recovery of activity was incomplete for TEM-1, K1 and P99 beta-lactamases, suggesting partitioning of the inhibited enzymes to give a permanently (or at least very stable) inactivated species. Examination of the interactions of the penem with TEM, B. cereus I and P99 beta-lactamases by e.s.m.s. also showed rapid and stoichiometric binding of the inhibitor. In all cases a mass increase of 264 Da over the native enzyme was observed, corresponding to the molecular mass of BRL 42715, showing that no fragmentation of the penem occurred on reaction with the beta-lactamases.

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Selected References

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

  1. Ambler R. P. The structure of beta-lactamases. Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):321–331. doi: 10.1098/rstb.1980.0049. [DOI] [PubMed] [Google Scholar]
  2. Aplin R. T., Baldwin J. E., Schofield C. J., Waley S. G. Use of electrospray mass spectrometry to directly observe an acyl enzyme intermediate in beta-lactamase catalysis. FEBS Lett. 1990 Dec 17;277(1-2):212–214. doi: 10.1016/0014-5793(90)80847-c. [DOI] [PubMed] [Google Scholar]
  3. Arisawa M., Then R. Inactivation of TEM-1 beta-lactamase by 6-acetylmethylenepenicillanic acid. Biochem J. 1983 Mar 1;209(3):609–615. doi: 10.1042/bj2090609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Aronoff S. C., Jacobs M. R., Johenning S., Yamabe S. Comparative activities of the beta-lactamase inhibitors YTR 830, sodium clavulanate, and sulbactam combined with amoxicillin or ampicillin. Antimicrob Agents Chemother. 1984 Oct;26(4):580–582. doi: 10.1128/aac.26.4.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barthélémy M., Peduzzi J., Labia R. Distinction entre les structures primaires des beta-lactamases TEM-1 et TEM-2. Ann Inst Pasteur Microbiol. 1985 May-Jun;136A(3):311–321. doi: 10.1016/s0769-2609(85)80093-4. [DOI] [PubMed] [Google Scholar]
  6. Bennett I. S., Brooks G., Broom N. J., Calvert S. H., Coleman K., François I. 6-(substituted methylene)penems, potent broad spectrum inhibitors of bacterial beta-lactamase. V. Chiral 1,2,3-triazolyl derivatives. J Antibiot (Tokyo) 1991 Sep;44(9):969–978. doi: 10.7164/antibiotics.44.969. [DOI] [PubMed] [Google Scholar]
  7. Bush K., Macalintal C., Rasmussen B. A., Lee V. J., Yang Y. Kinetic interactions of tazobactam with beta-lactamases from all major structural classes. Antimicrob Agents Chemother. 1993 Apr;37(4):851–858. doi: 10.1128/aac.37.4.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cartwright S. J., Waley S. G. beta-Lactamase inhibitors. Med Res Rev. 1983 Oct-Dec;3(4):341–382. doi: 10.1002/med.2610030402. [DOI] [PubMed] [Google Scholar]
  9. Charnas R. L., Knowles J. R. Inhibition of the RTEM beta-lactamase from Escherichia coli. Interaction of enzyme with derivatives of olivanic acid. Biochemistry. 1981 May 12;20(10):2732–2737. doi: 10.1021/bi00513a005. [DOI] [PubMed] [Google Scholar]
  10. Charnas R. L., Then R. L. Mechanism of inhibition of chromosomal beta-lactamases by third-generation cephalosporins. Rev Infect Dis. 1988 Jul-Aug;10(4):752–760. doi: 10.1093/clinids/10.4.752. [DOI] [PubMed] [Google Scholar]
  11. Coleman K., Griffin D. R., Page J. W., Upshon P. A. In vitro evaluation of BRL 42715, a novel beta-lactamase inhibitor. Antimicrob Agents Chemother. 1989 Sep;33(9):1580–1587. doi: 10.1128/aac.33.9.1580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. De Meester F., Matagne A., Dive G., Frère J. M. Unexpected influence of ionic strength on branched-pathway interactions between beta-lactamases and beta-halogenopenicillanates. Biochem J. 1989 Jan 1;257(1):245–249. doi: 10.1042/bj2570245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Easton C. J., Knowles J. R. Inhibition of the RTEM beta-lactamase from Escherichia coli. Interaction of the enzyme with derivatives of olivanic acid. Biochemistry. 1982 Jun 8;21(12):2857–2862. doi: 10.1021/bi00541a008. [DOI] [PubMed] [Google Scholar]
  14. Emanuel E. L., Gagnon J., Waley S. G. Structural and kinetic studies on beta-lactamase K1 from Klebsiella aerogenes. Biochem J. 1986 Mar 1;234(2):343–347. doi: 10.1042/bj2340343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. English A. R., Retsema J. A., Girard A. E., Lynch J. E., Barth W. E. CP-45,899, a beta-lactamase inhibitor that extends the antibacterial spectrum of beta-lactams: initial bacteriological characterization. Antimicrob Agents Chemother. 1978 Sep;14(3):414–419. doi: 10.1128/aac.14.3.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fisher J., Charnas R. L., Knowles J. R. Kinetic studies on the inactivation of Escherichia coli RTEM beta-lactamase by clavulanic acid. Biochemistry. 1978 May 30;17(11):2180–2184. doi: 10.1021/bi00604a024. [DOI] [PubMed] [Google Scholar]
  17. Frère J. M., Dormans C., Lenzini V. M., Duyckaerts C. Interaction of clavulanate with the beta-lactamases of Streptomyces albus G and Actinomadura R39. Biochem J. 1982 Dec 1;207(3):429–436. doi: 10.1042/bj2070429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Galleni M., Lindberg F., Normark S., Cole S., Honore N., Joris B., Frere J. M. Sequence and comparative analysis of three Enterobacter cloacae ampC beta-lactamase genes and their products. Biochem J. 1988 Mar 15;250(3):753–760. doi: 10.1042/bj2500753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hardy L. W., Kirsch J. F. Isolation of a Staphylococcus aureus beta-lactamase-dicloxacillin complex and kinetic studies on the reactivation of the enzyme. Arch Biochem Biophys. 1989 Jan;268(1):338–348. doi: 10.1016/0003-9861(89)90595-x. [DOI] [PubMed] [Google Scholar]
  20. Hunter P. A., Coleman K., Fisher J., Taylor D. In vitro synergistic properties of clavulanic acid, with ampicillin, amoxycillin and ticarcillin. J Antimicrob Chemother. 1980 Jul;6(4):455–470. doi: 10.1093/jac/6.4.455. [DOI] [PubMed] [Google Scholar]
  21. Jaurin B., Grundström T. ampC cephalosporinase of Escherichia coli K-12 has a different evolutionary origin from that of beta-lactamases of the penicillinase type. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4897–4901. doi: 10.1073/pnas.78.8.4897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. KITZ R., WILSON I. B. Esters of methanesulfonic acid as irreversible inhibitors of acetylcholinesterase. J Biol Chem. 1962 Oct;237:3245–3249. [PubMed] [Google Scholar]
  23. Madgwick P. J., Waley S. G. beta-lactamase I from Bacillus cereus. Structure and site-directed mutagenesis. Biochem J. 1987 Dec 15;248(3):657–662. doi: 10.1042/bj2480657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. O'Callaghan C. H., Morris A., Kirby S. M., Shingler A. H. Novel method for detection of beta-lactamases by using a chromogenic cephalosporin substrate. Antimicrob Agents Chemother. 1972 Apr;1(4):283–288. doi: 10.1128/aac.1.4.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Payne D. J., Skett P. W., Aplin R. T., Robinson C. V., Knowles D. J. beta-Lactamase ragged ends detected by electrospray mass spectrometry correlates poorly with multiple banding on isoelectric focusing. Biol Mass Spectrom. 1994 Mar;23(3):159–164. doi: 10.1002/bms.1200230307. [DOI] [PubMed] [Google Scholar]
  26. Reading C., Cole M. Clavulanic acid: a beta-lactamase-inhiting beta-lactam from Streptomyces clavuligerus. Antimicrob Agents Chemother. 1977 May;11(5):852–857. doi: 10.1128/aac.11.5.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reading C., Cole M. Structure-activity relationships amongst beta-lactamase inhibitors. J Enzyme Inhib. 1986;1(2):83–104. doi: 10.3109/14756368609020108. [DOI] [PubMed] [Google Scholar]
  28. Reading C., Farmer T. The inhibition of beta-lactamases from gram-negative bacteria by clavulanic acid. Biochem J. 1981 Dec 1;199(3):779–787. doi: 10.1042/bj1990779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Reading C., Hepburn P. The inhibition of staphylococcal beta-lactamase by clavulanic acid. Biochem J. 1979 Apr 1;179(1):67–76. doi: 10.1042/bj1790067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rizwi I., Tan A. K., Fink A. L., Virden R. Clavulanate inactivation of Staphylococcus aureus beta-lactamase. Biochem J. 1989 Feb 15;258(1):205–209. doi: 10.1042/bj2580205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Samuni A. A direct spectrophotometric assay and determination of Michaelis constants for the beta-lactamase reaction. Anal Biochem. 1975 Jan;63(1):17–26. doi: 10.1016/0003-2697(75)90185-2. [DOI] [PubMed] [Google Scholar]
  32. Waley S. G. A spectrophotometric assay of beta-lactamase action on penicillins. Biochem J. 1974 Jun;139(3):789–790. doi: 10.1042/bj1390789. [DOI] [PMC free article] [PubMed] [Google Scholar]

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