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

David M Mueller, Neeti Puri, Venkataraman Kabaleeswaran, Cassandra Terry, Andrew G W Leslie, John E Walker

Rosalind Franklin University of Medicine and Science, The Chicago Medical School, Department of Biochemistry and Molecular Biology, USA. david.mueller@rosalindfranklin.edu

Acta crystallographica. Section D, Biological crystallography 2004 Aug

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A genetically modified (His6-tagged) form of the mitochondrial F1-ATPase (MW = 370 kDa) has been purified from the yeast Saccharomyces cerevisiae and crystallized in the presence of polyethelene glycol (PEG) 6000 as a precipitant, 1 mM NiCl2, 1 mM Mg AMP-PNP and 50 microM 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 A. 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 A, beta = 101.6 degrees. The asymmetric unit contains three molecules of yeast F1, with a corresponding volume per protein weight (VM) of 2.8 A3 Da(-1) and a solvent content of 55%.

Citation

David M Mueller, Neeti Puri, Venkataraman Kabaleeswaran, Cassandra Terry, Andrew G W Leslie, John E Walker. Crystallization and preliminary crystallographic studies of the mitochondrial F1-ATPase from the yeast Saccharomyces cerevisiae. Acta crystallographica. Section D, Biological crystallography. 2004 Aug;60(Pt 8):1441-4