Open AccessX‐ray diffraction analysis of a crystal of HscA from Escherichia coli
Author(s) -
Aoto Phillip C.,
Ta Dennis T.,
CuppVickery Jill R.,
Vickery Larry E.
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105019251
Subject(s) - escherichia coli , x ray , diffraction , crystal (programming language) , crystallography , physics , materials science , chemistry , optics , computer science , biochemistry , programming language , gene
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 Å. The crystal belongs to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 158.35, b = 166.15, c = 168.26 Å, 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.