Altered penicillin-binding proteins in methicillin-resistant strains of Staphylococcus aureus.
The penicillin-binding proteins (PBPs) of a methicillin-resistant (MR) and a methicillin-susceptible (MS) Staphylococcus aureus were compared by various approaches involving the use of high-specific-activity [3H]penicillin as a reagent. The MR and MS strains were found to contain PBPs of the same number and electrophoretic mobilities. However, saturation of PBPs 1, 2, and 3 by methicillin in the MR strain required the use of several thousands of micrograms of antibiotic per milliliter, whereas 0.2 to 0.4 micrograms of methicillin per ml was sufficient to effectively compete with [3H]penicillin for the PBPs for the MS strain. Additional experiments indicate that these differences most likely reflect a greatly decreased affinity of the PBPs of the MR strain as compared to those of the MS strain. Shift of the pH of the culture medium of the MR strain from pH 7.0 to 5.2 resulted in an immediate drop in phenotypic resistance to methicillin (from a minimal inhibitory concentration value of 3,200 micrograms/ml at pH 7.0 to 0.8 microgram/ml at pH 5.2). Examination of the methicillin affinities of PBPs in MR bacteria grown at pH 5.2 showed the presence of the same low-affinity PBPs as in bacteria grown at pH 7.0. Thus, the pH-dependent resensitization to methicillin cannot be explained by a parallel increase in the antibiotic affinities of the PBPs.
D001426:Bacterial Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D003470:Culture Media; D006602:Hexosyltransferases; D006863:Hydrogen-Ion Concentration; D008712:Methicillin / Q000378:metabolism / Q000494:pharmacology*; D002267:Muramoylpentapeptide Carboxypeptidase; D009154:Mutation; D010400:Penicillin G / Q000378:metabolism; D010403:Penicillin Resistance; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003470:Culture Media; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D010400:Penicillin G; D008712:Methicillin