EXPRESSION AND CRYSTALLIZATION OF THE YEAST HSP82 CHAPERONE, AND PRELIMINARY-X-RAY DIFFRACTION STUDIES OF THE AMINO-TERMINAL DOMAIN

Expression of the Saccharomyces cerevisiae Hsp82 chaperone in a pep 4- 3-and hsc82-deficient strain of S. cerevisiae yielded over 25% of the total cell protein as intact Hsp82. Similarly, the amino-terminal doma in (residues 1-220) of Hsp82 was expressed to 18% of the total cell pr otein. Crystals of the intact Hsp82 were readily obtained, The crystal s were very fragile, suggesting a high solvent content, and diffracted to approximately 8 Angstrom. Tetragonal bipyrimidal crystals of the a mino-terminal domain of Hsp82 were readily obtained under a variety of different conditions. The crystals have primitive tetragonal space gr oup (P422, P4(1)22, or its enantiomorph P4(3)22) with unit cell dimens ions of a = 75.1 Angstrom and c 111.3 Angstrom, contain 60% by volume solvent, and diffract to 2.5 Angstrom resolution. Addition of 25% glyc erol to the mother liquor gave rise to large rod-shaped crystals. The crystals diffract to 2.8 Angstrom resolution, have an orthorhombic spa ce group (P222(1), P2(1)2(1)2, or P2(1)2(1)2(1)) with cell dimensions of a = 45.2 Angstrom, b = 115.4 Angstrom, and c = 116.9 Angstrom, and a solvent content of 58% by volume.