Purification, crystallization and preliminary X-ray diffraction studies of the PvuII endonuclease.
The PvuII endonuclease (PvuIIR) is a restriction enzyme from a type II restriction-modification system of Proteus vulgaris coded on plasmid pPvu1. The protein recognizes the DNA sequence 5' CAG'CTG 3' and shows no sequence homology to other restriction enzymes. This makes PvuIIR an interesting subject for structural determination. A purification procedure was developed that yields milligram quantities of the PvuIIR from plasmids expressed in the Escherichia coli strain HB101. The protein was crystallized using ammonium sulphate as precipitant. The crystals are orthorhombic, space group P2(1)2(1)2 with cell dimensions: a = 84.2 A, b = 106.2 A, c = 46.9 A. The asymmetric unit contains one PvuIIR dimer. Diffraction extends to 2.3 A, so the crystals may permit structural determination at atomic resolution.
Athanasiadis
A
A
University of Crete, Department of Biology, Greece.
Kokkinidis
M
M
D003461:Crystallography; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry* / Q000302:isolation & purification; D011514:Proteus vulgaris / Q000201:enzymology; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific