Vancomycin analogues active against vanA-resistant strains inhibit bacterial transglycosylase without binding substrate.
Bacterial transglycosylases are enzymes that couple the disaccharide subunits of peptidoglycan to form long carbohydrate chains. These enzymes are the target of the pentasaccharide antibiotic moenomycin as well as the proposed target of certain glycopeptides that overcome vancomycin resistance. Because bacterial transglycosylases are difficult enzymes to study, it has not previously been possible to evaluate how moenomycin inhibits them or to determine whether glycopeptide analogues directly target them. We have identified transglycosylase assay conditions that enable kinetic analysis of inhibitors and have examined the inhibition of Escherichia coli penicillin-binding protein 1b (PBP1b) by moenomycin as well as by various glycopeptides. We report that chlorobiphenyl vancomycin analogues that are incapable of binding substrates nevertheless inhibit E. coli PBP1b, which shows that these compounds interact directly with the enzyme. These findings support the hypothesis that chlorobiphenyl vancomycin derivatives overcome vanA resistance by targeting bacterial transglycosylases. We have also found that moenomycin is not competitive with respect to the lipid II substrate of PBP1b, as has long been believed. With the development of suitable methods to evaluate bacterial transglycosylases, it is now possible to probe the mechanism of action of some potentially very important antibiotics.
Chen
Lan
L
Department of Chemistry, Princeton University, Princeton, NJ 08544, USA.
Walker
Deborah
D
Sun
Binyuan
B
Hu
Yanan
Y
Walker
Suzanne
S
Kahne
Daniel
D
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins; D006602:Hexosyltransferases / Q000037:antagonists & inhibitors* / Q000235:genetics; D007700:Kinetics; D008968:Molecular Conformation; D015394:Molecular Structure; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors* / Q000235:genetics; D002267:Muramoylpentapeptide Carboxypeptidase; D016254:Mutagenesis, Insertional; D046915:Penicillin-Binding Proteins; D046928:Peptidoglycan Glycosyltransferase; D010458:Peptidyl Transferases / Q000037:antagonists & inhibitors* / Q000235:genetics; D046929:Serine-Type D-Ala-D-Ala Carboxypeptidase; D014640:Vancomycin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D020713:Vancomycin Resistance / Q000502:physiology*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003062:Codon; D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D046915:Penicillin-Binding Proteins; D014640:Vancomycin; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D046928:Peptidoglycan Glycosyltransferase; C478681:penicillin-binding protein 1B, E coli; D046929:Serine-Type D-Ala-D-Ala Carboxypeptidase; D002267:Muramoylpentapeptide Carboxypeptidase