Sg. Li et al., PURIFICATION AND CRYSTAL-GROWTH OF F1-ATPASE FROM PIG-HEART MITOCHONDRIA, Biochemistry and molecular biology international, 40(3), 1996, pp. 479-486
A method has been evolved toward the aim of getting suitable crystals
for high resolution of structural analysis of F-1-ATPase by X-ray crys
tallography. The different conditions for crystal growth of ATPase tha
t were isolated and purified by different methods from pig heart mitoc
hondrial ATP synthase had been compared and screened. A simple method
for purification of F-1-ATPase was adopted. Tile F-1-ATPase is release
d with chloroform from submitochondrial particles. Then it was treated
with fractional precipitation of (NH4)(2)SO4 and finally was further
purified by employing the sephadex G 200 column. The crystals of F-1-A
TPase were usually obtained after a few mounths. They appeared to have
uniform morphology of tetrahedron. They diffracted to a resolution of
7 Angstrom. The diffraction data were collected on the XRD-100 Siemen
s Area Detector. According to a total of 240 frames, the cell paramete
rs obtained are a = b = 147 Angstrom, c = 208 Angstrom, alpha = beta =
gamma = 90 degrees, tile probable space group is P4 or its antipode.
The reproducibility of this method for crystallization of FI-ATPase is
good.