Crystallization and preliminary crystallographic analysis of deoxyuridine 5 '-triphosphate nucleotidohydrolase from Bacillus subtilis

Single crystals of purified homotrimeric deoxyuridine 5'-triphosphate nucle otidohydrolase (dUTPase) from Bacillus subtilis have been grown under sever al different conditions using vapour diffusion. X-ray diffraction data have been collected using synchrotron radiation from three crystal forms of the unliganded enzyme and from enzyme cocrystallized with a substrate analogue and inhibitor, dUDP, and a metal ion, Sr2+. The three crystal forms of unl iganded enzyme belong to hexagonal (P6(3)), orthorhombic (P2(1)2(1)2) and c ubic (P2(1)3) space groups and data have been recorded to 1.75, 1.90 and 2. 50 Angstrom spacing, respectively. Crystals grown in the presence of dUDP a nd Sr2+ belong to the orthorhombic space group P2(1)2(1)2(1) and data were measured to 1.90 Angstrom spacing. Solution of the hexagonal crystal form b y molecular replacement using the dUTPase from feline immunodeficiency viru s as a search model is in progress.