Crystallization and preliminary X-ray structural analysis of nucleoside triphosphate hydrolases from Neospora caninum and Toxoplasma gondii

Kazuaki Matoba, Tomoo Shiba, Tsutomu Takeuchi, L. David Sibley, Makiko Seiki, Fumi Kikyo, Toshio Horiuchi, Takashi Asai, Shigeharu Harada

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The nucleoside triphosphate hydrolases that are produced by Neospora caninum (NcNTPase) and Toxoplasma gondii (TgNTPase-I) have a different physio-logical function from the ubiquitous ecto-ATPases. The recombinant enzymes were crystallized at 293 K using polyethylene glycol 3350 as a precipitant and X - ray diffraction data sets were collected for NcNTPase (to 2.8 Å resolution) and TgNTPase-I (to 3.1 Å resolution) at 100 K using synchrotron radiation. The crystals of NcNTPase and TgNTPase-I belonged to the orthorhombic space group I222 (unit-cell parameters a = 93.6, b = 140.8, c = 301.1 Å) and the monoclinic space group P21 (unit-cell parameters a = 87.1, b = 123.5, c = 120.2 Å, β = 96.6°), respectively, with two NcNTPase (V M = 3.7 Å3 Da-1) and four TgNTPase-I (V M = 2.7 Å3 Da-1) molecules per asymmetric unit. SAD phasing trials using a data set (λ = 0.97904 Å) collected from a crystal of seleno-methionylated NcNTPase gave an initial electron-density map of sufficient quality to build a molecular model of NcNTPase.

Original languageEnglish
Pages (from-to)1445-1448
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume66
Issue number11
DOIs
Publication statusPublished - 2010 Nov

Fingerprint

N-Glycosyl Hydrolases
Neospora
nucleosides
Toxoplasma
Hydrolases
Crystallization
Nucleosides
structural analysis
Structural analysis
X-Rays
crystallization
X rays
Crystals
Synchrotrons
Molecular Models
Synchrotron radiation
cells
X-Ray Diffraction
crystals
Carrier concentration

Keywords

  • Neospora caninum
  • nucleoside triphosphate hydrolases
  • Toxoplasma gondii

ASJC Scopus subject areas

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

Cite this

Crystallization and preliminary X-ray structural analysis of nucleoside triphosphate hydrolases from Neospora caninum and Toxoplasma gondii. / Matoba, Kazuaki; Shiba, Tomoo; Takeuchi, Tsutomu; Sibley, L. David; Seiki, Makiko; Kikyo, Fumi; Horiuchi, Toshio; Asai, Takashi; Harada, Shigeharu.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 66, No. 11, 11.2010, p. 1445-1448.

Research output: Contribution to journalArticle

Matoba, Kazuaki ; Shiba, Tomoo ; Takeuchi, Tsutomu ; Sibley, L. David ; Seiki, Makiko ; Kikyo, Fumi ; Horiuchi, Toshio ; Asai, Takashi ; Harada, Shigeharu. / Crystallization and preliminary X-ray structural analysis of nucleoside triphosphate hydrolases from Neospora caninum and Toxoplasma gondii. In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications. 2010 ; Vol. 66, No. 11. pp. 1445-1448.
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