Crystallization of Chlorella deoxyuridine triphosphatase

Laura Badalucco, Ishwari Poudel, Mamoru Yamanishi, Chandrasekhar Natarajan, Hideaki Moriyama

Research output: Contribution to journalArticle

3 Citations (Scopus)

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′-[(,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, = 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.

Original languageEnglish (US)
Pages (from-to)1599-1602
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume67
Issue number12
DOIs
StatePublished - Dec 1 2011

Fingerprint

Chlorella
Deoxyuridine
Crystallization
crystallization
Genes
genes
enzymes
Ammonium Sulfate
Synthetic Genes
Enzymes
Algae
Manganese
ammonium sulfates
Needles
Magnesium
Escherichia coli
algae
Escherichia
Codon
X-Ray Diffraction

Keywords

  • Chlorella
  • Chlorella virus
  • dUTPases

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

Cite this

Crystallization of Chlorella deoxyuridine triphosphatase. / Badalucco, Laura; Poudel, Ishwari; Yamanishi, Mamoru; Natarajan, Chandrasekhar; Moriyama, Hideaki.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 67, No. 12, 01.12.2011, p. 1599-1602.

Research output: Contribution to journalArticle

Badalucco, Laura ; Poudel, Ishwari ; Yamanishi, Mamoru ; Natarajan, Chandrasekhar ; Moriyama, Hideaki. / Crystallization of Chlorella deoxyuridine triphosphatase. In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications. 2011 ; Vol. 67, No. 12. pp. 1599-1602.
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