Crystallization of the flexible nuclear import receptor importin-beta in the unliganded state

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Jun 1;65(Pt 6):625-8. doi: 10.1107/S1744309109016820. Epub 2009 May 22.

Abstract

The transport of macromolecules across the nuclear envelope is an essential eukaryotic process that enables proteins such as transcription factors, polymerases and histones to gain access to the genetic material contained within the nucleus. Importin-beta plays a central role in the nucleocytoplasmic transport process, mediating nuclear import through a range of interactions with cytoplasmic, nuclear and nuclear pore proteins such as importin-alpha, Ran, nucleoporins and various cargo molecules. The unliganded form of the full-length yeast importin-beta has been expressed and crystallized. The crystals were obtained by vapour diffusion at pH 6.5 and 290 K. The crystals belonged to space group P2(1) (unit-cell parameters a = 58.17, b = 127.25, c = 68.52 A, beta = 102.23). One molecule is expected in the asymmetric unit. The crystals diffracted to 2.4 A resolution using a laboratory X-ray source and were suitable for crystal structure determination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus
  • Amino Acid Sequence
  • Cell Nucleus / metabolism*
  • Crystallization
  • Data Collection
  • Escherichia coli / genetics
  • Glutathione Transferase / metabolism
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Particle Size
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Scattering, Small Angle
  • Solubility
  • Statistics as Topic
  • Temperature
  • Transformation, Bacterial
  • X-Ray Diffraction
  • beta Karyopherins / chemistry
  • beta Karyopherins / isolation & purification
  • beta Karyopherins / metabolism*

Substances

  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • beta Karyopherins
  • Glutathione Transferase