Protein preparation, crystallization and preliminary X-ray analysis of imidazolonepropionase from Bacillus subtilis

Biochim Biophys Acta. 2006 Jan;1764(1):153-6. doi: 10.1016/j.bbapap.2005.08.012. Epub 2005 Sep 12.

Abstract

Imidazolonepropionase (EC 3.5.2.7) is the third enzyme of the histidine degradation pathway that has been conserved from bacteria to eukaryotes. The enzyme is the only one with unknown three-dimensional structure in this pathway. In this work, Bacillus subtilis imidazolonepropionase (HutI) was expressed in E. coli and purified to homogeneity. After thrombin digestion, high quality crystals were obtained by hanging-drop vapor diffusion method. The best crystal diffracted to 2.0 A and belonged to the space group P2(1) with unit-cell parameters a = 57.73 A, b = 106.34 A, c = 66.47 A, beta = 89.93 degrees .

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / chemistry
  • Amidohydrolases / genetics
  • Amidohydrolases / isolation & purification*
  • Amidohydrolases / metabolism
  • Bacillus subtilis / enzymology*
  • Bacillus subtilis / genetics
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Bacterial Proteins / metabolism
  • Crystallization
  • Crystallography, X-Ray
  • Genes, Bacterial
  • Histidine / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • Histidine
  • Amidohydrolases
  • imidazolonepropionase, Bacillus subtilis