Open AccessCrystallization and preliminary X‐ray structural analysis of nucleoside triphosphate hydrolases from Neospora caninum and Toxoplasma gondii
Author(s) -
Matoba Kazuaki,
Shiba Tomoo,
Takeuchi Tsutomu,
Sibley L. David,
Seiki Makiko,
Kikyo Fumi,
Horiuchi Toshio,
Asai Takashi,
Harada Shigeharu
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110032136
Subject(s) - neospora caninum , orthorhombic crystal system , crystallography , toxoplasma gondii , monoclinic crystal system , polyethylene glycol , crystallization , chemistry , resolution (logic) , molecule , crystal structure , biology , biochemistry , antibody , organic chemistry , immunology , artificial intelligence , computer science
The nucleoside triphosphate hydrolases that are produced by Neospora caninum (NcNTPase) and Toxoplasma gondii (TgNTPase‐I) have a different physiological 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 I 222 (unit‐cell parameters a = 93.6, b = 140.8, c = 301.1 Å) and the monoclinic space group P 2 1 (unit‐cell parameters a = 87.1, b = 123.5, c = 120.2 Å, β = 96.6°), respectively, with two NcNTPase ( V M = 3.7 Å 3 Da −1 ) and four TgNTPase‐I ( V M = 2.7 Å 3 Da −1 ) molecules per asymmetric unit. SAD phasing trials using a data set (λ = 0.97904 Å) collected from a crystal of selenomethionylated NcNTPase gave an initial electron‐density map of sufficient quality to build a molecular model of NcNTPase.