Open AccessCrystallization and preliminary crystallographic studies of both components of the staphylococcal LukE–LukD leukotoxin
Author(s) -
Galy Romain,
Bergeret Fabien,
Keller Daniel,
Mourey Lionel,
Prévost Gilles,
Maveyraud Laurent
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112014662
Subject(s) - crystallization , crystallography , molecular replacement , protein data bank (rcsb pdb) , chemistry , resolution (logic) , recombinant dna , escherichia coli , diffraction , molecule , crystal structure , stereochemistry , physics , biochemistry , optics , organic chemistry , artificial intelligence , computer science , gene
Soluble forms of recombinant LukE protein (expressed in Escherichia coli ) and of wild‐type LukD protein (expressed in Staphylococcus aureus ), which together form the staphylococcal LukE–LukD leukotoxin, were purified to homogeneity and crystallized using the sitting‐drop vapour‐diffusion method. The crystals of LukE belonged to space group I 4, with unit‐cell parameters a = b = 134.50, c = 64.43 Å, and diffracted X‐rays to 1.6 Å resolution. The crystals of LukD belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 48.04, b = 50.99, c = 137.40 Å, and diffracted to 1.9 Å resolution. Molecular replacement using the LukF‐PV structure (PDB entry 1pvl ) as a template model allowed the identification of an initial structure solution for the LukD data. In the case of LukE, a solution comprising only a single copy of the search model (LukS‐PV; PDB entry 1t5r ) was found, although the unit‐cell parameters indicated that up to three molecules could be accommodated in the asymmetric unit.