title: Crystallization of Chlorella deoxyuridine triphosphatase.
abstract: Deoxyuridine triphosphatase (dUTPase) is a ubiquitous enzyme that has been widely studied owing to its function and evolutionary significance. The gene coding for the dUTPase from the Chlorella alga was codon-optimized and synthesized. The synthetic gene was expressed in Escherichia coli and recombinant core Chlorella dUTPase (chdUTPase) was purified. Crystallization of chdUTPase was performed by the repetitive hanging-drop vapor-diffusion method at 298 K with ammonium sulfate as the precipitant. In the presence of 2'-deoxyuridine-5'-[(α,β)-imido]triphosphate and magnesium, the enzyme produced die-shaped hexagonal R3 crystals with unit-cell parameters a = b = 66.9, c = 93.6 Å, γ = 120°. X-ray diffraction data for chdUTPase were collected to 1.6 Å resolution. The crystallization of chdUTPase with manganese resulted in very fragile clusters of needles.
authors.affiliation: School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68588-0118, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Badalucco
authors.forename: Ishwari
authors.initials: I
authors.lastname: Poudel
authors.forename: Mamoru
authors.initials: M
authors.lastname: Yamanishi
authors.forename: Chandrasekhar
authors.initials: C
authors.lastname: Natarajan
authors.forename: Hideaki
authors.initials: H
authors.lastname: Moriyama
mesh_terms: D002708:Chlorella / Q000201:enzymology*; D003460:Crystallization; D011755:Pyrophosphatases / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase