Crystallization and preliminary crystallographic studies of both components of the staphylococcal LukE-LukD leukotoxin.
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 I4, 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 P2(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.
Institut de Pharmacologie et de Biologie Structurale, Centre National de la Recherche Scientifique, 205 Route de Narbonne, BP 64182, F-31077 Toulouse, France.
Romain
R
Galy
Fabien
F
Bergeret
Daniel
D
Keller
Lionel
L
Mourey
Gilles
G
Prévost
Laurent
L
Maveyraud
D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D005098:Exotoxins / Q000737:chemistry*; D013211:Staphylococcus aureus / Q000737:chemistry*
D001426:Bacterial Proteins; D005098:Exotoxins; C115218:LukD protein, Staphylococcus aureus; C115217:LukE protein, Staphylococcus aureus