Protein preparation, crystallization and preliminary X-ray crystallographic studies of a thermostable hypoxanthine-guanine phosphoribosyltransferase from Thermoanaerobacter tengcongensis

Acta Crystallogr D Biol Crystallogr. 2003 Oct;59(Pt 10):1863-5. doi: 10.1107/s0907444903018250. Epub 2003 Sep 19.

Abstract

Native and His-tagged mutant (L160I) hypoxanthine-guanine phosphoribosyltransferase (HGPRT) from Thermoanaerobacter tengcongensis were cloned, expressed in Escherichia coli and purified. Both proteins were crystallized with polyethylene glycol as the main precipitant at 293 K using the hanging-drop vapour-diffusion method. The crystal of native HGPRT belongs to space group C222(1), with unit-cell parameters a = 65.77, b = 137.73, c = 95.27 A, and diffracted to 2.2 A resolution on an in-house X-ray generator. The crystal of the His-tagged mutant (L160I) HGPRT belongs to the space group I222, with unit-cell parameters a = 52.21, b = 88.36, c = 93.03 A, and diffracted to 1.7 A resolution in-house.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillaceae / enzymology*
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Crystallization / methods
  • Crystallography, X-Ray
  • Enzyme Stability
  • Hypoxanthine Phosphoribosyltransferase / biosynthesis*
  • Hypoxanthine Phosphoribosyltransferase / chemistry*
  • Hypoxanthine Phosphoribosyltransferase / genetics
  • Protein Conformation
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Temperature

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • Hypoxanthine Phosphoribosyltransferase