Cubic crystals of chloramphenicol phosphotransferase from Streptomyces venezuelae in complex with chloramphenicol.
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.
Ellis
J
J
Department of Biochemistry, University of Leicester, Leicester LE1 7RH, England.
Campopiano
D J
DJ
Izard
T
T
D001426:Bacterial Proteins; D002701:Chloramphenicol / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry; D013302:Streptomyces / Q000201:enzymology*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D002701:Chloramphenicol; D017853:Phosphotransferases (Alcohol Group Acceptor); C096858:chloramphenicol 3'-O-phosphotransferase