Crystallization of Chlorella deoxyuridine triphosphatase

Authors

Laura Badalucco, University of Nebraska-Lincoln
Ishwari Poudel, University of Nebraska-LincolnFollow
Mamoru Yamanishi, University of Nebraska-Lincoln
Chandrasekhar Natarajan, University of Nebraska-LincolnFollow
Hideaki Moriyama, University of Nebraska-LincolnFollow

Document Type

Article

Date of this Version

2011

Citation

Acta Cryst. (2011). F67, 1599–1602

doi:10.1107/S1744309111038097

Comments

Copyright 2011 International Union of Crystallography All rights reserved

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 A, ƴ = 120°. X-ray diffraction data for chdUTPase were collected to 1.6 A resolution. The crystallization of chdUTPase with manganese resulted in very fragile clusters of needles.