title: X-ray diffraction analysis of a crystal of HscA from Escherichia coli.
abstract: HscA is a constitutively expressed Hsp70 that interacts with the iron-sulfur cluster assembly protein IscU. Crystals of a truncated form of HscA (52 kDa; residues 17-505) grown in the presence of an IscU-recognition peptide, WELPPVKI, have been obtained by hanging-drop vapor diffusion using ammonium sulfate as the precipitant. A complete native X-ray diffraction data set was collected from a single crystal at 100 K to a resolution of 2.9 A. The crystal belongs to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 158.35, b = 166.15, c = 168.26 A, and contains six molecules per asymmetric unit. Phases were determined by molecular replacement using the nucleotide-binding domain from DnaK and the substrate-binding domain from HscA as models. This is the first reported crystallization of an Hsp70 containing both nucleotide- and substrate-binding domains.
authors.affiliation: Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA.
authors.forename: Phillip C
authors.initials: PC
authors.lastname: Aoto
authors.forename: Dennis T
authors.initials: DT
authors.lastname: Ta
authors.forename: Jill R
authors.initials: JR
authors.lastname: Cupp-Vickery
authors.forename: Larry E
authors.initials: LE
authors.lastname: Vickery
mesh_terms: D000645:Ammonium Sulfate / Q000494:pharmacology; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry*; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry; D009711:Nucleotides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D009711:Nucleotides; D011994:Recombinant Proteins; C085650:hscA protein, E coli; C063842:dnaK protein, E coli; D000645:Ammonium Sulfate