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 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.