Abstract
The prfA genes of Bacillus stearothermophilus and Bacillus subtilis are in an operon downstream of the ponA gene encoding penicillin-binding protein 1 (PBP1), a major enzyme involved in peptidoglycan synthesis. The specific function of the 23- to 24-kDa PrfA protein is unknown but this protein plays some role in nucleoid segregation and the functions of PrfA and PBP1 are interrelated. We overexpressed B. stearothermophilus and B. subtilis PrfA in Escherichia coli and purified the proteins to homogeneity by cation exchange and gel filtration chromatography. The protein is a monomer in solution, and circular dichroism spectroscopy revealed an abundance of beta-sheet secondary structure. Crystals of B. stearothermophilus PrfA were also obtained and diffracted X-rays to 1.8 A resolution.
Copyright 2000 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Bacillus subtilis / chemistry*
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Bacillus subtilis / genetics
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Bacterial Proteins / chemistry*
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Bacterial Proteins / genetics
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Bacterial Proteins / isolation & purification
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Carrier Proteins / chemistry*
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Circular Dichroism
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Crystallization
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Crystallography, X-Ray
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Geobacillus stearothermophilus / chemistry*
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Geobacillus stearothermophilus / genetics
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Hexosyltransferases*
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Muramoylpentapeptide Carboxypeptidase / chemistry*
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Penicillin-Binding Proteins
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Peptidyl Transferases*
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Protein Structure, Secondary
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Solutions
Substances
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Bacterial Proteins
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Carrier Proteins
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Penicillin-Binding Proteins
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Recombinant Proteins
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Solutions
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Peptidyl Transferases
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Hexosyltransferases
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Muramoylpentapeptide Carboxypeptidase