Crystallization and Initial X-Ray Diffraction Characterization of Complexes of FxFG Nucleoporin Repeats with Nuclear Transport Factors

Autor: Bayliss, Richard, Kent, Helen M., Corbett, Anita H., Stewart, Murray
Zdroj: Journal of Structural Biology; September 2000, Vol. 131 Issue: 3 p240-247, 8p
Abstrakt: NTF2 and importin-β are transport factors that mediate nuclear protein import and which interact with nuclear pore proteins (nucleoporins) during translocation from the cytoplasm to the nucleus through nuclear pore complexes. We employed a native gel electrophoresis method to assess the interaction of nucleoporin constructs that contain FxFG sequence repeats with NTF2 and truncation mutants of importin-β to determine suitable fragments for crystallization. Based on these data, we obtained crystals of complexes between yeast NTF2 and a construct containing five FxFG nucleoporin repeats from the yeast nucleoporin Nsp1p and between a construct containing residues 1–442 of human importin-β and the same nucleoporin construct. The yeast NTF2–nucleoporin crystals have trigonal symmetry and diffract past 2.8 Å resolution using synchrotron radiation, whereas the importin-β–nucleoporin complex crystals have P21212 orthorhombic symmetry and diffract past 3.2 Å resolution.
Databáze: Supplemental Index
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