Crystallization and preliminary X-ray crystallographic analysis of p24, a component of the potato nuclear factor PBF-2

被引:5
作者
Desveaux, D [1 ]
Allard, J [1 ]
Brisson, N [1 ]
Sygusch, J [1 ]
机构
[1] Univ Montreal, Dept Biochim, Montreal, PQ H3C 3J7, Canada
来源
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY | 2002年 / 58卷
关键词
D O I
10.1107/S0907444901018741
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 Angstrom 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) Angstrom. The asymmetric unit contains four protomers, giving a crystal volume per protein mass (V-M) of 2.23 Angstrom(3) Da(-1) and a solvent content of 45% by volume.
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页码:296 / 298
页数:3
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