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

Acta Crystallogr D Biol Crystallogr. 2002 Feb;58(Pt 2):296-8. doi: 10.1107/s0907444901018741. Epub 2002 Jan 24.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Crystallization
  • Crystallography, X-Ray
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / metabolism
  • Plant Proteins / chemistry*
  • Protein Conformation
  • Selenomethionine / chemistry
  • Solanum tuberosum / chemistry*

Substances

  • DNA-Binding Proteins
  • Nuclear Proteins
  • PBF-2 protein, Solanum tuberosum
  • Plant Proteins
  • Selenomethionine