Expression, purification, crystallization and preliminary crystallographic analysis of human Rad GTPase

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Nov 1;61(Pt 11):978-80. doi: 10.1107/S1744309105031982. Epub 2005 Oct 20.

Abstract

Human Rad is a new member of the Ras GTPase superfamily and is overexpressed in human skeletal muscle of individuals with type II diabetes. The GTPase core domain was overexpressed in Escherichia coli and purified for crystallization. Crystals were obtained at 293 K by vapour diffusion using a crystallization robot. The crystals were found to belong to space group P2(1), with unit-cell parameters a = 52.2, b = 58.6, c = 53.4 A, beta = 97.9 degrees , and contained two Rad molecules in the crystallographic asymmetric unit. A diffraction data set was collected to a resolution of 1.8 A using synchrotron radiation at SPring-8.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray
  • Dithiothreitol / chemistry
  • Escherichia coli / metabolism
  • GTP Phosphohydrolases / chemistry
  • Humans
  • Models, Molecular
  • Protein Conformation
  • Synchrotrons
  • Temperature
  • Tissue Distribution
  • X-Ray Diffraction
  • ras Proteins / biosynthesis
  • ras Proteins / chemistry*
  • ras Proteins / isolation & purification

Substances

  • RRAD protein, human
  • GTP Phosphohydrolases
  • ras Proteins
  • Dithiothreitol