Abstract
Chloramphenicol 3-O-phosphotransferase (CPT) from Streptomyces venezuelae ISP5230, a novel chloramphenicol-inactivating kinase, has been overexpressed and purified using Escherichia coli as the heterologous host. Crystals of CPT in complex with its substrate chloramphenicol (Cm) were obtained which were suitable for X-ray diffraction. The crystals belong to the cubic space group I4132 with unit-cell dimension a = 200.0 A. The initial CPT crystals diffracted to 3.5 A and the diffraction was improved significantly upon adding acetonitrile and Cm to the crystallization drop. The CPT-Cm crystals diffract to at least 2.8 A resolution.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Bacterial Proteins*
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Chloramphenicol / chemistry*
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Crystallization
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Crystallography, X-Ray
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Phosphotransferases (Alcohol Group Acceptor) / chemistry*
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Phosphotransferases (Alcohol Group Acceptor) / genetics
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Phosphotransferases (Alcohol Group Acceptor) / isolation & purification
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Recombinant Proteins / chemistry
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Streptomyces / enzymology*
Substances
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Bacterial Proteins
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Recombinant Proteins
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Chloramphenicol
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Phosphotransferases (Alcohol Group Acceptor)
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chloramphenicol 3'-O-phosphotransferase