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. 1984 Sep;26(3):335–338. doi: 10.1128/aac.26.3.335

Affinity of temocillin for Escherichia coli K-12 penicillin-binding proteins.

R Labia, P Baron, J M Masson, G Hill, M Cole
PMCID: PMC176164  PMID: 6391369

Abstract

Temocillin is a 6 alpha-methoxy penicillin which shows poor affinity for the penicillin-binding proteins (PBPs) of Escherichia coli K-12 when tested by competition with [14C]penicillin G or [125I]penicillin X. When the reaction conditions for the radiolabeled penicillin used in this procedure were modified by lowering the temperature (2 degrees C) and reducing the incubation time (3 min), temocillin showed a much higher affinity for PBP-3 and improved affinity for the other PBPs, with the exception of PBP-2. Direct labeling procedures with [14C]temocillin also showed that the compound had affinity for PBPs 1a and 3. These results are more consistent with the effects of temocillin on the morphology of E. coli than the poor affinity values obtained by the classical competitive procedure. Reasons for the disparity between these assay systems are discussed.

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Selected References

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  1. Berenguer J., De Pedro M. A., Vázquez D. V. Interaction of nocardicin A with the penicillin-binding proteins of Escherichia coli in intact cells and in purified cell envelopes. Eur J Biochem. 1982 Aug;126(1):155–159. doi: 10.1111/j.1432-1033.1982.tb06760.x. [DOI] [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Coyette J., Ghuysen J. M., Fontana R. Solubilization and isolation of the membrane-bound DD-carboxypeptidase of Streptococcus faecalis ATCC9790. Properties of the purified enzyme. Eur J Biochem. 1978 Jul 17;88(1):297–305. doi: 10.1111/j.1432-1033.1978.tb12450.x. [DOI] [PubMed] [Google Scholar]
  4. Curtis N. A., Brown C., Boxall M., Boulton M. G. Inhibition of Escherichia coli K-12 by beta-lactam antibiotics with poor antibacterial activity: interaction of permeability and intrinsic activity against penicillin-binding proteins. Antimicrob Agents Chemother. 1979 Mar;15(3):332–336. doi: 10.1128/aac.15.3.332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Curtis N. A., Orr D., Ross G. W., Boulton M. G. Affinities of penicillins and cephalosporins for the penicillin-binding proteins of Escherichia coli K-12 and their antibacterial activity. Antimicrob Agents Chemother. 1979 Nov;16(5):533–539. doi: 10.1128/aac.16.5.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Curtis N. A., Ross G. W., Boulton M. G. Effect of 7-alpha methoxy substitution of cephalosporins upon their affinity for the penicillin-binding proteins of E. coli K12. Comparison with antibacterial activity and inhibition of membrane bound model transpeptidase activity. J Antimicrob Chemother. 1979 Jul;5(4):391–398. doi: 10.1093/jac/5.4.391. [DOI] [PubMed] [Google Scholar]
  7. Filip C., Fletcher G., Wulff J. L., Earhart C. F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. doi: 10.1128/jb.115.3.717-722.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Imada A., Kitano K., Kintaka K., Muroi M., Asai M. Sulfazecin and isosulfazecin, novel beta-lactam antibiotics of bacterial origin. Nature. 1981 Feb 12;289(5798):590–591. doi: 10.1038/289590a0. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  11. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  12. Masson J. M., Labia R. Synthesis of a 125I-radiolabeled penicillin for penicillin-binding proteins studies. Anal Biochem. 1983 Jan;128(1):164–168. doi: 10.1016/0003-2697(83)90357-3. [DOI] [PubMed] [Google Scholar]
  13. Ohya S., Yamazaki M., Sugawara S., Tamaki S., Matsuhashi M. New cephamycin antibiotic, CS-1170: binding affinity to penicillin-binding proteins and inhibition of peptidoglycan cross-linking reactions in Escherichia coli. Antimicrob Agents Chemother. 1978 Nov;14(5):780–785. doi: 10.1128/aac.14.5.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schilf W., Frère P., Frère J. M., Martin H. H., Ghuysen J. M., Adriaens P., Meesschaert B. Interaction between penicillin and the DD-carboxypeptidase of the unstable L-form of Proteus mirabilis strain 19. Eur J Biochem. 1978 Apr 17;85(2):325–330. doi: 10.1111/j.1432-1033.1978.tb12242.x. [DOI] [PubMed] [Google Scholar]
  15. Slocombe B., Basker M. J., Bentley P. H., Clayton J. P., Cole M., Comber K. R., Dixon R. A., Edmondson R. A., Jackson D., Merrikin D. J. BRL 17421, a novel beta-lactam antibiotic, highly resistant to beta-lactamases, giving high and prolonged serum levels in humans. Antimicrob Agents Chemother. 1981 Jul;20(1):38–46. doi: 10.1128/aac.20.1.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Spratt B. G., Jobanputra V. Mutants of Escherichia coli which lack a component of penicillin-binding protein 1 are viable. FEBS Lett. 1977 Jul 15;79(2):374–378. doi: 10.1016/0014-5793(77)80824-7. [DOI] [PubMed] [Google Scholar]
  17. Spratt B. G., Pardee A. B. Penicillin-binding proteins and cell shape in E. coli. Nature. 1975 Apr 10;254(5500):516–517. doi: 10.1038/254516a0. [DOI] [PubMed] [Google Scholar]
  18. Spratt B. G. Properties of the penicillin-binding proteins of Escherichia coli K12,. Eur J Biochem. 1977 Jan;72(2):341–352. doi: 10.1111/j.1432-1033.1977.tb11258.x. [DOI] [PubMed] [Google Scholar]
  19. Umbreit J. N., Strominger J. L. D-alanine carboxypeptidase from Bacillus subtilis membranes. II. Interaction with penicillins and cephalosporins. J Biol Chem. 1973 Oct 10;248(19):6767–6771. [PubMed] [Google Scholar]
  20. Yourassowsky E., Van der Linden M. P., Lismont M. J., Crokaert F. Growth curve patterns and bacterial morphology of Escherichia coli subjected to different temocillin (BRL 17421) concentrations. J Antimicrob Chemother. 1982 Oct;10(4):289–293. doi: 10.1093/jac/10.4.289. [DOI] [PubMed] [Google Scholar]

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