Abstract
The mecA-27r gene, which encodes PBP2a-27r, was modified by site-specific mutagenesis, resulting in replacement of the N-terminal membrane anchor with a short chelating peptide (CP-PBP2a-27r). CP-PBP2a-27r retained the same binding affinity for beta-lactam antibiotics as the wild-type enzyme. Approximately 95% pure CP-PBP2a-27r was recovered in a single step by use of chelating-peptide-immobilized metal ion affinity chromatography.
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins*
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Base Sequence
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Carrier Proteins / biosynthesis*
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Carrier Proteins / isolation & purification*
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Carrier Proteins / metabolism
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Cefamandole / metabolism
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Chromatography, Affinity
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DNA Primers
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Genes, Bacterial
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Hexosyltransferases*
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Methicillin Resistance / genetics
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Methicillin Resistance / physiology*
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Molecular Sequence Data
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Muramoylpentapeptide Carboxypeptidase / biosynthesis*
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Muramoylpentapeptide Carboxypeptidase / isolation & purification*
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Muramoylpentapeptide Carboxypeptidase / metabolism
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Mutagenesis, Site-Directed
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Penicillin-Binding Proteins
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Peptidyl Transferases*
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Promoter Regions, Genetic
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Staphylococcus aureus / genetics
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Staphylococcus aureus / metabolism*
Substances
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Bacterial Proteins
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Carrier Proteins
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DNA Primers
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Penicillin-Binding Proteins
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Recombinant Proteins
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Cefamandole
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Peptidyl Transferases
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Hexosyltransferases
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Muramoylpentapeptide Carboxypeptidase