Expression, purification and crystallization of a functional core of the voltage-dependent calcium channel beta subunit

Acta Crystallogr D Biol Crystallogr. 2004 Jul;60(Pt 7):1301-3. doi: 10.1107/S0907444904010686. Epub 2004 Jun 22.

Abstract

Two versions of the functional core of the rabbit voltage-dependent calcium channel beta2a subunit were expressed in Escherichia coli. These proteins were purified to homogeneity and screened for crystallization. Crystallization conditions were refined using the hanging-drop vapour-diffusion method and two crystal forms were pursued. Crystal form I is represented by thick rods with tetragonal symmetry, unit-cell parameters a = b = 75, c = 165 A and a diffraction limit of 3.4 A which were obtained using ammonium sulfate as a precipitant. Crystal form II gives rise to plates with orthorhombic symmetry, unit-cell parameters a = 35, b = 75, c = 165 A and a diffraction limit of 2.3 A which were grown using polyethylene glycol 20K as a precipitant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium Channels / chemistry*
  • Calcium Channels / genetics
  • Calcium Channels / isolation & purification*
  • Crystallization
  • Escherichia coli / genetics
  • Gene Expression
  • Protein Subunits / chemistry*
  • Protein Subunits / genetics
  • Protein Subunits / isolation & purification*

Substances

  • Calcium Channels
  • Protein Subunits