Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1981 Oct 1;199(1):129–136. doi: 10.1042/bj1990129

Interaction between non-classical beta-lactam compounds and the Zn2+-containing G and serine R61 and R39 D-alanyl-D-alanine peptidases.

J A Kelly, J M Frère, D Klein, J M Ghuysen
PMCID: PMC1163342  PMID: 6279094

Abstract

Streptomyces albus G secretes a Zn2+-containing D-alanyl-D-alanine peptidase. Streptomyces R61 and Actinomadura R39 secrete D-alanyl-D-alanine-cleaving serine peptidases. The effect of non-classical beta-lactam antibiotics on these three model enzymes has been studied. Mecillinam, cefoxitin, quinacillin, quinacillin sulphone, clavulanate and N-formimidoylthienamycin have no effect on the Zn2+-containing enzyme. 6-Amino-penicillanic acid slowly inactivates this enzyme and 7-aminocephalosporanic acid behaves as a reversible inhibitor. Cefoxitin and N-formimidoylthienamycin are potent anti-bacterial agents; they effectively inactivate the serine R39 enzyme and, to a lesser extent, the serine R61 enzyme. All the other beta-lactam compounds tested, including mecillinam, are slow inactivators of these serine enzymes. The intermediates formed between 6-aminopenicillanic acid and the R61 and R39 enzymes are long- and short-lived respectively, whereas those formed between 7-aminocephalosporanic acid and the same R61 and R39 enzymes are short- and long-lived respectively. Breakdown of the short-lived intermediates thus obtained gives rise to several ninhydrin-positive degradation products. The intermediates formed between clavulanate and the serine enzymes are long-lived. With the R39 enzyme, the inactivated complex formed in a first step undergoes subsequent monomolecular rearrangement to give rise to a second species exhibiting a high absorbance at 273 nm.

Full text

PDF
136

Selected References

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

  1. Cartwright S. J., Coulson A. F. A semi-synthetic penicillinase inactivator. Nature. 1979 Mar 22;278(5702):360–361. doi: 10.1038/278360a0. [DOI] [PubMed] [Google Scholar]
  2. Dideberg O., Charlier P., Dupont L., Vermeire M., Frere J. M., Ghuysen J. M. The 4.5 A resolution structure analysis of the exocellular DD-carboxypeptidase of Streptomyces albus G. FEBS Lett. 1980 Aug 11;117(1):212–214. doi: 10.1016/0014-5793(80)80947-1. [DOI] [PubMed] [Google Scholar]
  3. Dideberg O., Joris B., Frere J. M., Ghuysen J. M., Weber G., Robaye R., Delbrouck J. M., Roelandts I. The exocellular DD-carboxypeptidase of Streptomyces albus G: a metallo (Zn2+) enzyme. FEBS Lett. 1980 Aug 11;117(1):215–218. doi: 10.1016/0014-5793(80)80948-3. [DOI] [PubMed] [Google Scholar]
  4. Duez C., Frère J. M., Geurts F., Ghuysen J. M., Dierickx L., Delcambe L. The exocellular DD-carboxypeptidase-endopeptidase from Streptomyces albus G. Purification and chemical properties. Biochem J. 1978 Dec 1;175(3):793–800. doi: 10.1042/bj1750793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Duez C., Frère J. M., Klein D., Noël M., Ghuysen J. M., Delcambe L., Dierickx L. The exocellular beta-lactamase of Streptomyces albus G. Purification, properties and comparison with the exocellular DD-carboxypeptidase. Biochem J. 1981 Jan 1;193(1):75–82. doi: 10.1042/bj1930075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duez C., Joris B., Frère J. M., Ghuysen J. M., Van Beeumen J. The penicillin-binding site in the exocellular DD-carboxypeptidase-transpeptidase of Actinomadura R39. Biochem J. 1981 Jan 1;193(1):83–86. doi: 10.1042/bj1930083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fisher J., Belasco J. G., Charnas R. L., Khosla S., Knowles J. R. Beta-lactamase inactivation by mechanism-based reagents. Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):309–319. doi: 10.1098/rstb.1980.0048. [DOI] [PubMed] [Google Scholar]
  8. Frere J., Ghuysen J., Degelaen J., Loffet A., Perkins H. R. Fragmentation of benzylpenicillin after interaction with the exocellular DD-carboxypeptidase-transpeptidases of Streptomyces R61 and R39. Nature. 1975 Nov 13;258(5531):168–170. doi: 10.1038/258168a0. [DOI] [PubMed] [Google Scholar]
  9. Frieden C. Kinetic aspects of regulation of metabolic processes. The hysteretic enzyme concept. J Biol Chem. 1970 Nov 10;245(21):5788–5799. [PubMed] [Google Scholar]
  10. Frère J. M., Duez C., Ghuysen J. M., Vandekerkhove J. Occurrence of a serine residue in the penicillin-binding site of the exocellular DD-carboxy-peptidase-transpeptidase from Streptomyces R61. FEBS Lett. 1976 Nov;70(1):257–260. doi: 10.1016/0014-5793(76)80770-3. [DOI] [PubMed] [Google Scholar]
  11. Frère J. M., Geurts F., Ghuysen J. M. The exocellular DD-carboxypeptidase-endopeptidase of Streptomyces albus G. Interaction with beta-lactam antibiotics. Biochem J. 1978 Dec 1;175(3):801–805. doi: 10.1042/bj1750801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Frère J. M., Ghuysen J. M., Iwatsubo M. Kinetics of interaction between the exocellular DD-carboxypeptidase-transpeptidase from Streptomyces R61 and beta-lactam antibiotics. A choice of models. Eur J Biochem. 1975 Sep 15;57(2):343–351. doi: 10.1111/j.1432-1033.1975.tb02307.x. [DOI] [PubMed] [Google Scholar]
  13. Frère J. M., Ghuysen J. M., Perkins H. R., Nieto M. Molecular weight and amino acid composition of the exocellular DD-carboxypeptidase-transpeptidase of Streptomyces R61. Biochem J. 1973 Nov;135(3):463–468. doi: 10.1042/bj1350463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Frère J. M., Moreno R., Ghuysen J. M. Molecular weight, amino acid composition and physicochemical properties of the exocellular DD-carboxypeptidase-transpeptidase of Streptomyces R39. Biochem J. 1974 Oct;143(1):233–240. doi: 10.1042/bj1430233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fuad N., Frère J. M., Ghuysen J. M., Duez C., Iwatsubo M. Mode of interaction between beta-lactam antibiotics and the exocellular DD-carboxypeptidase--transpeptidase from Streptomyces R39. Biochem J. 1976 Jun 1;155(3):623–629. doi: 10.1042/bj1550623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ghuysen J. M., Frère J. M., Leyh-Bouille M., Coyette J., Dusart J., Nguyen-Distèche M. Use of model enzymes in the determination of the mode of action of penicillins and delta 3-cephalosporins. Annu Rev Biochem. 1979;48:73–101. doi: 10.1146/annurev.bi.48.070179.000445. [DOI] [PubMed] [Google Scholar]
  17. Johnson K., Duez C., Frère J. M., Ghuysen J. M. Beta-lactamases (Actinomycetes species). Methods Enzymol. 1975;43:687–698. doi: 10.1016/0076-6879(75)43134-2. [DOI] [PubMed] [Google Scholar]
  18. Nieto M., Perkins H. R., Frère J. M., Ghuysen J. M. Fluorescence and circular dichroism studies on the Streptomyces R61 DD-carboxypeptidase-transpeptidase. Penicillin binding by the enzyme. Biochem J. 1973 Nov;135(3):493–505. doi: 10.1042/bj1350493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Callaghan C. H. Description and classification of the newer cephalosporins and their relationships with the established compounds. J Antimicrob Chemother. 1979 Nov;5(6):635–671. doi: 10.1093/jac/5.6.635. [DOI] [PubMed] [Google Scholar]
  20. Perkins H. R., Nieto M., Frére J. M., Leyh-Bouille M., Ghuysen J. M. Streptomyces DD-carboxypeptidases as transpeptidases. The specificity for amino compounds acting as carboxyl acceptors. Biochem J. 1973 Apr;131(4):707–718. doi: 10.1042/bj1310707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Thatcher D. R. The partial amino acid sequence of the extracellular beta-lactamase I of Bacillus cereus 569/H. Biochem J. 1975 May;147(2):313–326. doi: 10.1042/bj1470313. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES