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

Single crystals of purified homotrimeric deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) from Bacillus subtilis have been grown under several 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 unliganded enzyme belong to hexagonal (P6₃), orthorhombic (P2₁2₁2) and cubic (P2₁3) space groups and data have been recorded to 1.75, 1.90 and 2.50 Å spacing, respectively. Crystals grown in the presence of dUDP and Sr²⁺ belong to the orthorhombic space group P2₁2₁2₁ and data were measured to 1.90 Å spacing. Solution of the hexagonal crystal form by molecular replacement using the dUTPase from feline immunodeficiency virus as a search model is in progress.