Crystallization and X-ray data analysis of the 10 kDa C-terminal lid subdomain from Caenorhabditis elegans Hsp70.
Hsp70 is an important molecular chaperone involved in the regulation of protein folding. Crystals of the C-terminal 10 kDa helical lid domain (residues 542-640) from a Caenorhabditis elegans Hsp70 homologue have been produced that diffract X-rays to approximately 3.4 A. Crystals belong to space group I2(1)2(1)2(1), with unit-cell parameters a = b = 197, c = 200 A. The Matthews coefficient, self-rotation function and Patterson map indicate 24 monomers in the asymmetric unit, showing non-crystallographic 432 symmetry. Molecular-replacement studies using the corresponding domain from rat, the only eukaryotic homologue with a known structure, failed and a mercury derivative was obtained. Preliminary MAD phasing using SHELXD and SHARP for location and refinement of the heavy-atom substructure and SOLOMON for density modification produced interpretable maps with a clear protein-solvent boundary. Further density-modification, model-building and refinement are currently under way.
Worrall
Liam
L
School of Biological Sciences, University of Edinburgh, The King's Buildings, Mayfield Road, Edinburgh EH9 3JR, Scotland.
Walkinshaw
Malcolm D
MD
D000818:Animals; D017173:Caenorhabditis elegans / Q000737:chemistry*; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification; D017434:Protein Structure, Tertiary
D029742:Caenorhabditis elegans Proteins; D018840:HSP70 Heat-Shock Proteins; D010446:Peptide Fragments