Crystallization and crystal-packing studies of Chlorella virus deoxyuridine triphosphatase

Kohei Homma, Hideaki Moriyama

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

The 141-amino-acid deoxyuridine triphosphatase (dUTPase) from the algal Chlorella virus IL-3A and its Glu81Ser/Thr84Arg-mutant derivative Mu-22 were crystallized using the hanging-drop vapor-diffusion method at 298 K with polyethylene glycol as the precipitant. An apo IL-3A dUTPase with an amino-terminal T7 epitope tag and a carboxy-terminal histidine tag yielded cubic P213 crystals with unit-cell parameter a = 106.65 Å. In the presence of dUDP, the enzyme produced thin stacked orthorhombic P222 crystals with unit-cell parameters a = 81.0, b = 96.2, c = 132.8 Å. T7-histidine-tagged Mu-22 dUTPase formed thin stacked rectangular crystals. Amino-terminal histidine-tagged dUTPases did not crystallize but formed aggregates. Glycyl-seryl-tagged dUTPases yielded cubic P213 IL-3A crystals with unit-cell parameter a = 105.68 Å and hexagonal P63 Mu-22 crystals with unit-cell parameters a = 132.07, c = 53.45 Å, = 120°. Owing to the Thr84Arg mutation, Mu-22 dUTPase had different monomer-to-monomer interactions to those of IL-3A dUTPase.

Original languageEnglish (US)
Pages (from-to)1030-1034
Number of pages5
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume65
Issue number10
DOIs
StatePublished - 2009

Keywords

  • Chlorella
  • Chlorella virus
  • DUTPases

ASJC Scopus subject areas

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

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