Article (Scientific journals)
Membrane-bound transpeptidase and penicillin binding sites in Streptomyces strain R61
Marquet, Alberto; Dusart, Jean; Ghuysen, Jean-Marie et al.
1974In European Journal of Biochemistry, 46 (3), p. 515-523
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Keywords :
acyltransferases metabolism; ampicillin/pharmacology; binding sites; binding, competitive; carbenicillin/pharmacology; cell membrane/enzymology; cephalosporins/metabolism/pharmacology; chromatography, thin layer; cloxacillin/pharmacology; electrophoresis, paper; kinetics; methicillin/pharmacology; oxacillin/pharmacology; penicillin g/*metabolism/pharmacology; penicillin v/pharmacology; penicillinase/metabolism; peptidoglycan/biosynthesis; protein binding; streptomyces/*metabolism; sulfhydryl reagents/pharmacology; transferases/antagonists & inhibitors
Abstract :
[en] High-affinity penicillin binding sites from which the antibiotic could not be removed by washings at 4°C in 0.017 M K2HPO4 or 0.05 M Tris-HCl pH 7.5, were shown to occur in the isolated membranes of Streptomyces R61. These sites caused the attachment of 25 picomoles of [14C]benzylpenicillin per milligram membrane protein. Penicillins and cephalosporins competed for the same binding sites. The antibiotic concentrations which excluded [14C]benzylpenicillin from 50% of the binding sites were those which inhibited by 50% the membrane-bound transpeptidase. The same rate constant (about 1 × 10−4 s−1) for the dissociation of the benzylpenicillin membrane complex at 37°C and in 0.017 M K2HPO4, was calculated either from the release of the radioactivity (using [14C]benzylpenicillin) or from the recovery of the transpeptidase activity. These observations supported the conclusion that the high-affinity binding sites in the isolated membranes were the transpeptidase molecules. All the complexes formed between the membranes and the various penicillins and cephalosporins examined were reversible at 37°C and in 0.017 M K2HPO4 at least with regard to the transpeptidase. Depending upon the antibiotics, the rate constants for the dissociation of these complexes varied from 3.3 × 10−3 to 0.73 × 10−4 s−1. The radioactivity released through the dissociation of [14C]benzylpenicillin membrane complex occurred mainly in the form of a compound which behaved as [W]-benzylpenicilloic acid both by paper electrophoresis and thin-layer chromatography. It was impossible to choose between several possible mechanisms for the release of the antibiotic molecule.
Disciplines :
Biochemistry, biophysics & molecular biology
Microbiology
Author, co-author :
Marquet, Alberto;  Université de Liège - ULiège > Faculté de Médecine, Institut de Botanique > Service de Microbiologie
Dusart, Jean;  Université de Liège - ULiège > Faculté de Médecine, Institut de Botanique > Service de Microbiologie
Ghuysen, Jean-Marie ;  Université de Liège - ULiège > Faculté de Médecine, Institut de Botanique > Service de Microbiologie
Perkins, Harnold-R.;  National Institute for Medical Research
Language :
English
Title :
Membrane-bound transpeptidase and penicillin binding sites in Streptomyces strain R61
Publication date :
01 August 1974
Journal title :
European Journal of Biochemistry
ISSN :
0014-2956
eISSN :
1432-1033
Publisher :
Blackwell Science, Oxford, United Kingdom
Volume :
46
Issue :
3
Pages :
515-523
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
FRFC - Fonds de la Recherche Fondamentale Collective [BE]
Available on ORBi :
since 28 January 2011

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