Interaction of monoclonal antibodies with the enzymatic domains of penicillin-binding protein 1b of Escherichia coli.
Monoclonal antibodies (MAbs) against four different antigenic determinants of penicillin-binding protein (PBP) 1b were used to study the transglycosylase and transpeptidase activities of PBP 1b. Enzyme kinetics in the presence of and without the MAbs were determined, and the synthesized murein was analyzed. Two MAbs against the transglycosylase domain of PBP 1b appeared to inhibit this reaction. One MAb inhibited only the transpeptidase reaction, and one inhibited both enzymatic activities of PBP 1b. The latter two MAbs bound to the transpeptidase domain of PBP 1b. The following major conclusions were deduced from the results. (i) Transpeptidation is the rate-limiting step of the reaction cascade, and it is dependent on the product of transglycosylation. (ii) PBP 1b has only one type of transpeptidase activity, i.e., a penta-tetra transpeptidase activity. (iii) PBP 1b is probably a globular protein which has two intimately associated enzymatic domains.
Department of Molecular Cell Biology, University of Amsterdam, The Netherlands.
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den Blaauwen
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Aarsman
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Nanninga
D000217:Acyltransferases / Q000032:analysis* / Q000037:antagonists & inhibitors*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004926:Escherichia coli / Q000032:analysis*; D006602:Hexosyltransferases / Q000032:analysis* / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D007700:Kinetics; D009097:Multienzyme Complexes / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D002267:Muramoylpentapeptide Carboxypeptidase; D010400:Penicillin G / Q000378:metabolism; D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan / Q000096:biosynthesis; D010458:Peptidyl Transferases / Q000032:analysis* / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D002352:Carrier Proteins; D009097:Multienzyme Complexes; D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan; D000217:Acyltransferases; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D010400:Penicillin G