Open AccessCrystallization of Chlorella deoxyuridine triphosphatase
Author(s) -
Badalucco Laura,
Poudel Ishwari,
Yamanishi Mamoru,
Natarajan Chandrasekhar,
Moriyama Hideaki
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111038097
Subject(s) - crystallization , ammonium sulfate , chemistry , deoxyuridine , enzyme , escherichia coli , biochemistry , crystallography , gene , dna , chromatography , organic chemistry
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 R 3 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.