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 language | English |
|---|---|
| Pages (from-to) | 1445-1448 |
| Number of pages | 4 |
| Journal | Acta Crystallographica Section F: Structural Biology and Crystallization Communications |
| Volume | 66 |
| Issue number | 11 |
| DOIs | |
| Publication status | Published - 2010 Nov |
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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 journal › Article
}
TY - JOUR
T1 - Crystallization and preliminary X-ray structural analysis of nucleoside triphosphate hydrolases from Neospora caninum and Toxoplasma gondii
AU - Matoba, Kazuaki
AU - Shiba, Tomoo
AU - Takeuchi, Tsutomu
AU - Sibley, L. David
AU - Seiki, Makiko
AU - Kikyo, Fumi
AU - Horiuchi, Toshio
AU - Asai, Takashi
AU - Harada, Shigeharu
PY - 2010/11
Y1 - 2010/11
N2 - 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.
AB - 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.
KW - Neospora caninum
KW - nucleoside triphosphate hydrolases
KW - Toxoplasma gondii
UR - http://www.scopus.com/inward/record.url?scp=78149300135&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78149300135&partnerID=8YFLogxK
U2 - 10.1107/S1744309110032136
DO - 10.1107/S1744309110032136
M3 - Article
C2 - 21045291
AN - SCOPUS:78149300135
VL - 66
SP - 1445
EP - 1448
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
SN - 1744-3091
IS - 11
ER -