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Purification and crystal growth of F1‐ATPase from pig heart mitochondria
Author(s) -
Li ShengGuang,
Gui LuLu,
Lin Zhihuan,
Wan ZhuLi,
Chang WenRui,
Liang DongCai
Publication year - 1996
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549600201043
Subject(s) - atpase , crystallization , sephadex , atp synthase , chromatography , chloroform , chemistry , resolution (logic) , crystallography , enzyme , biochemistry , organic chemistry , artificial intelligence , computer science
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 7Å. 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=147Å, c=208Å, α=β=γ=90°, the probable space group is P4 or its antipode. The reproducibility of this method for crystallization of F1‐ATPase is good.