Cloning and characterization of a gene, pbpF, encoding a new penicillin-binding protein, PBP2B, in Staphylococcus aureus

A previously unrecognized penicillin binding protein (PBP) gene, pbpF, was identified in Staphylococcus aureus. This gene encodes a protein of 691 amino acid residues with an estimated molecular mass of 78 kDa. The molecular mass is very close to that of S. aureus PBP2 (81 kDa), and the protein is t...

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Bibliographic Details
Published in:Antimicrobial agents and chemotherapy 1999-07, Vol.43 (7), p.1578-1583
Main Authors: KOMATSUZAWA, H, CHOI, G. H, OHTA, K, SUGAI, M, TRAN, M. T, SUGINAKA, H
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacterial Proteins
Bacteriology
Base Sequence
Biological and medical sciences
Carrier Proteins - chemistry
Carrier Proteins - genetics
Carrier Proteins - immunology
Cloning, Molecular
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genetics
Hexosyltransferases
Mechanisms of Resistance
Microbiology
Molecular Sequence Data
Muramoylpentapeptide Carboxypeptidase - chemistry
Muramoylpentapeptide Carboxypeptidase - genetics
Muramoylpentapeptide Carboxypeptidase - immunology
Penicillin G - metabolism
Penicillin-Binding Proteins
Peptidyl Transferases
Precipitin Tests
Staphylococcus aureus
Staphylococcus aureus - genetics
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Summary:A previously unrecognized penicillin binding protein (PBP) gene, pbpF, was identified in Staphylococcus aureus. This gene encodes a protein of 691 amino acid residues with an estimated molecular mass of 78 kDa. The molecular mass is very close to that of S. aureus PBP2 (81 kDa), and the protein is tentatively named PBP2B. PBP2B has three motifs, SSVK, SSN, and KTG, that can be found in PBPs and beta-lactamases. Recombinant PBP2B (rPBP2B), which lacks a putative signal peptide at the N terminus and has a histidine tag at the C terminus, was expressed in Escherichia coli. The purified rPBP2B was shown to have penicillin binding activity. A protein band was detected from S. aureus membrane fraction by immunoblotting with anti-rPBP2B serum. Also, penicillin binding activity of the protein immunoprecipitated with anti-rPBP2B serum was detected. These results suggest the presence of PBP2B in S. aureus cell membrane that covalently binds penicillin. The internal region of pbpF and PBP2B protein were found in all 12 S. aureus strains tested by PCR and immunoblotting.
ISSN:0066-4804
1098-6596
DOI:10.1128/AAC.43.7.1578