Abstract
The high-molecular-mass penicillin-binding proteins (HMM-PBPs), present in the cytoplasmic membranes of all eubacteria, are involved in important physiological events such as cell elongation, septation or shape determination. Up to now it has, however, been very difficult or impossible to study the catalytic properties of the HMM-PBPs in vitro. With simple substrates, we could demonstrate that several of these proteins could catalyse the hydrolysis of some thioesters or the transfer of their acyl moiety on the amino group of a suitable acceptor nucleophile. Many of the acyl-donor substrates were hippuric acid or benzoyl-D-alanine derivatives, and their spectroscopic properties enabled a direct monitoring of the enzymic reaction. In their presence, the binding of radioactive penicillin to the PBPs was also inhibited.
MeSH Terms
Acyltransferases/metabolism
Aminobutyrates/metabolism
Bacterial Proteins
Carrier Proteins/metabolism
Catalysis
Cell Membrane/chemistry
Enterococcus/chemistry
Escherichia coli/chemistry
Esters/metabolism,pharmacology
Hexosyltransferases
Hippurates/metabolism
Hydrolysis
Kinetics
Molecular Weight
Muramoylpentapeptide Carboxypeptidase/metabolism
Penicillin G/metabolism
Penicillin-Binding Proteins
Peptidyl Transferases
Streptomyces/chemistry
Substrate Specificity
Sulfhydryl Compounds/metabolism,pharmacology
Chemicals
Aminobutyrates
Bacterial Proteins
Carrier Proteins
Esters
Hippurates
Penicillin-Binding Proteins
Sulfhydryl Compounds
2-amino-4-phenylbutyric acid
Acyltransferases
Peptidyl Transferases
Hexosyltransferases
Muramoylpentapeptide Carboxypeptidase
Penicillin G
hippuric acid
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Adam M
Centre d'Ingénierie des Protéines, Université de Liège, Belgium.
Damblon C
Jamin M
Zorzi W
Dusart V
Galleni M
el Kharroubi A
Piras G
Spratt B G
Keck W
References (13)
13 references, click to expand
-
Enzymic mechanisms involving concomitant transfer and hydrolysis reactions.
Biochem J. 1973 Nov;135(3):469-81
PMID: 4772273
-
Penicillin-binding proteins and cell shape in E. coli.
Nature. 1975 Apr 10;254(5500):516-7
PMID: 1091862
-
Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12.
Proc Natl Acad Sci U S A. 1975 Aug;72(8):2999-3003
PMID: 1103132
-
Properties of the penicillin-binding proteins of Escherichia coli K12,.
Eur J Biochem. 1977 Jan;72(2):341-52
PMID: 319999
-
In vitro peptidoglycan polymerization catalysed by penicillin binding protein 1b of Escherichia coli K-12.
FEBS Lett. 1980 Feb 11;110(2):245-9
PMID: 6989636
-
On the process of cellular division in Escherichia coli: isolation and characterization of penicillin-binding proteins 1a, 1b, and 3.
Proc Natl Acad Sci U S A. 1980 Aug;77(8):4499-503
PMID: 7001458
-
Serine beta-lactamases and penicillin-binding proteins.
Annu Rev Microbiol. 1991;45:37-67
PMID: 1741619
-
Streptomyces K15 DD-peptidase-catalysed reactions with ester and amide carbonyl donors.
Biochem J. 1986 Apr 1;235(1):167-76
PMID: 2874789
-
Automated analysis of enzyme inactivation phenomena. Application to beta-lactamases and DD-peptidases.
Biochem Pharmacol. 1987 Jul 15;36(14):2393-403
PMID: 3038122
-
Overexpression, solubilization and refolding of a genetically engineered derivative of the penicillin-binding protein 3 of Escherichia coli K12.
Mol Microbiol. 1988 Jul;2(4):519-25
PMID: 3050360
-
Chromogenic depsipeptide substrates for beta-lactamases and penicillin-sensitive DD-peptidases.
Biochem J. 1990 Sep 1;270(2):525-9
PMID: 2400398
-
Characterization of an Enterococcus hirae penicillin-binding protein 3 with low penicillin affinity.
J Bacteriol. 1990 Dec;172(12):6856-62
PMID: 2254261
-
Penicillin-sensitive enzymes in peptidoglycan biosynthesis.
Crit Rev Microbiol. 1985;11(4):299-396
PMID: 3888533