Crystallization and preliminary crystallographic studies of the mitochondrial F1-ATPase from the yeast Saccharomyces cerevisiae

A genetically modified (His(6)-tagged) form of the mitochondrial F-1-ATPase ( MW = 370 kDa) has been purified from the yeast Saccharomyces cerevisiae and crystallized in the presence of polythelene glycol ( PEG) 6000 as a precipitant, 1 mM NiCl2, 1 mM Mg AMP-PNP and 50 muM Mg ADP. X-ray diffraction data were obtained on three separate occasions using synchrotron radiation, with a progression in the quality of the diffraction data, which improved from 3.3 to 3.0 to 2.8 Angstrom. On the second occasion, the diffraction was improved by a crystal-annealing procedure. The crystals belong to the monoclinic space group P2(1), with unit-cell parameters a = 110.6, b = 294.2, c = 190.4 Angstrom, beta = 101.6degrees. The asymmetric unit contains three molecules of yeast F-1, with a corresponding volume per protein weight (V-M) of 2.8 Angstrom(3) Da(-1) and a solvent content of 55%.