Expression, crystallization and phasing of vacuolar H+-ATPase subunit C (Vma5p) of Saccharomyces cerevisiae

The expression, crystallization and phasing of subunit C (Vma5p) of the yeast ( Saccharomyces cerevisiae) vacuolar proton-translocating ATPase (V-ATPase) is described. The expressed protein consists of 412 residues: 392 from the reading frame of Vma5p and 20 N-terminal residues originating from the plasmid. Diffraction-quality crystals were obtained using the hanging-drop and sitting-drop vapour-diffusion methods assisted by streak-seeding, with PEG 3350 as precipitant. The crystals formed in hanging drops diffracted to 1.80 Angstrom and belong to space group P4(3)2(1)2(1), with unit-cell parameters a = b = 62.54, c = 327.37 Angstrom, alpha = beta = gamma = 90degrees. The structure was solved using SIRAS with a Lu(O2C2H3)(2) heavy-atom derivative.