FemA of Staphylococcus aureus: isolation and immunodetection.
FemA, a cytoplasmic protein necessary for the expression of methicillin resistance in Staphylococcus aureus and also involved in the biosynthesis of staphylococcal cell walls, was detected and quantified in several S. aureus strains under different growth conditions by Western immunoblot. Two types of antigens were used for the production of polyclonal antibodies against FemA: (i) a synthetic peptide comprising 14 amino acids of its C-terminal sequence; and (ii) FemA isolated by preparative gel electrophoresis and electroelution from an overproducing staphylococcal strain. Immunodetection revealed that all investigated strains, either methicillin-resistant or susceptible, expressed FemA during the exponential growth phase in varying amounts. In the stationary phase, the FemA content was diminished. Strains in which femA was inactivated by insertion of Tn551 into the control region of the femAB operon still expressed about 10% of the protein compared to their parent strains. Tn551 insertion in the middle of the femB gene did not affect the FemA expression. In 40 methicillin-susceptible and 6 resistant clinical isolates of S. aureus, the FemA content or its affinity to the antibodies was reduced compared to laboratory parent strains. In susceptible strains, an additional protein of higher molecular weight, present in large quantities, was also able to bind the FemA antibodies. Such a protein was also present in methicillin-resistant isolates, although it was not as pronounced as in the susceptible strains.
Johnson
S
S
Robert Koch-Institute, Berlin, Germany.
Krüger
D
D
Labischinski
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H
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D016106:Methicillin Resistance; D010446:Peptide Fragments / Q000276:immunology; D013045:Species Specificity; D013211:Staphylococcus aureus / Q000737:chemistry* / Q000276:immunology
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; C096686:FemA protein, Bacteria; D010446:Peptide Fragments