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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'-[(?,?)-imido]triphosphate and magnesium, the enzyme produced die-shaped hexagonal R3 crystals with unit-cell parameters a = b = 66.9, c = 93.6 Å, ? = 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.

SUBMITTER: Badalucco L

PROVIDER: S-EPMC3232149 | biostudies-literature | 2011 Dec

REPOSITORIES: biostudies-literature

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Crystallization of Chlorella deoxyuridine triphosphatase.

Badalucco Laura L Poudel Ishwari I Yamanishi Mamoru M Natarajan Chandrasekhar C Moriyama Hideaki H

Acta crystallographica. Section F, Structural biology and crystallization communications 20111126 Pt 12

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 ...[more]

PMID: 22139176

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