Crystallization and preliminary X-ray crystallographic analysis of p24, a component of the potato nuclear factor PBF-2.
The Solanum tuberosum (potato) nuclear factor PBF-2 is implicated in pathogen-induced expression of the pathogenesis-related gene PR-10a. Crystals of the DNA-binding component of PBF-2, p24, have been obtained at 277 K in 20 mM Tris-HCl pH 8.0. Recombinant protein with a His tag at its C-terminus was overexpressed in Escherichia coli in the presence and absence of selenomethionine and was purified using a combination of HiTrap affinity columns and gel-filtration chromatography. Crystals suitable for structural analysis were obtained for both native and selenomethionine-labelled proteins and yielded diffraction data at 100 K that were processed to 2.3 and 2.8 A resolution, respectively. The p24 protein crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 69.4 (69.1), b = 89.4 (90.5), c = 144.1 (144.3) A. The asymmetric unit contains four protomers, giving a crystal volume per protein mass (V(M)) of 2.23 A(3) Da(-1) and a solvent content of 45% by volume.
Desveaux
Darrell
D
Département de Biochimie, Université de Montréal, CP 6128, Station Centre Ville, Montréal, Québec H3C 3J7, Canada.
Allard
Julie
J
Brisson
Normand
N
Sygusch
Jurgen
J
D003460:Crystallization; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry*; D011487:Protein Conformation; D012645:Selenomethionine / Q000737:chemistry; D011198:Solanum tuberosum / Q000737:chemistry*
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C416516:PBF-2 protein, Solanum tuberosum; D010940:Plant Proteins; D012645:Selenomethionine