Purification and crystal growth of F1-ATPase from pig heart mitochondria.
A method has been evolved toward the aim of getting suitable crystals for high resolution of structural analysis of F1-ATPase by X-ray crystallography. The different conditions for crystal growth of ATPase that were isolated and purified by different methods from pig heart mitochondrial ATP synthase had been compared and screened. A simple method for purification of F1-ATPase was adopted. The F1-ATPase is released with chloroform from submitochondrial particles. Then it was treated with fractional precipitation of (NH4)2SO4 and finally was further purified by employing the sephadex G 200 column. The crystals of F1-ATPase were usually obtained after a few months. They appeared to have uniform morphology of tetrahedron. They diffracted to a resolution of 7A. The diffraction data were collected on the XRD-100 Siemens Area Detector. According to a total of 240 frames, the cell parameters obtained are a = b = 147 A, c = 208 A, alpha = beta = gamma = 90 alpha, the probable space group is P4 or its antipode. The reproducibility of this method for crystallization of F1-ATPase is good.
Li
S G
SG
Institute of Biophysics, Chinese Academy of Sciences, National Laboratory of Biomacromolecules, Beijing, China.
Gui
L L
LL
Lin
Z H
ZH
Wan
Z L
ZL
Chang
W R
WR
Liang
D C
DC
D000818:Animals; D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D008929:Mitochondria, Heart / Q000737:chemistry / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000302:isolation & purification; D013552:Swine; D014961:X-Ray Diffraction
D006180:Proton-Translocating ATPases