Expression, purification, crystallization and preliminary X-ray diffraction analysis of uracil phosphoribosyltransferase of Toxoplasma gondii

Acta Crystallogr D Biol Crystallogr. 1999 Jan;55(Pt 1):347-9. doi: 10.1107/S090744499800821X. Epub 1999 Jan 1.

Abstract

Recombinant uracil phosphoribosyltransferase (UPRT) enzyme of Toxoplasma gondii was expressed in Escherichia coli and purified from the cell-free extract by a combination of chromatographic steps. The recombinant protein was enzymatically active when tested in an in vitro UPRT assay. The purified protein was crystallized using the hanging-drop vapor-diffusion technique with ammonium phosphate as precipitant. The crystallized protein also exhibited UPRT activity. Crystals diffract to 2.4 A resolution and belong to space group P3121 or P3221 with unit-cell dimensions a = b = 119.9, c = 70.8 A and two molecules per asymmetric unit.

Publication types

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

MeSH terms

  • Animals
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • Gene Expression
  • Pentosyltransferases / chemistry*
  • Pentosyltransferases / genetics
  • Pentosyltransferases / isolation & purification*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Toxoplasma / enzymology*
  • Toxoplasma / genetics

Substances

  • Recombinant Proteins
  • Pentosyltransferases
  • uracil phosphoribosyltransferase