Crystallization and preliminary X-ray studies of a recombinant calcium-binding protein from Entamoeba histolytica

Acta Crystallogr D Biol Crystallogr. 1998 Nov 1;54(Pt 6 Pt 2):1442-5. doi: 10.1107/s0907444998001759.

Abstract

A calcium-binding protein (CaBP) of Entamoeba histolytica was purified from an E. coli recombinant clone carrying the CaBP gene in a pET-3c expression vector using anion-exchange and size-exclusion chromatography. Examination of the amino-acid sequence of the recombinant protein suggested that it has four independent EF-hand motifs. The protein dissolved in cacodylate buffer was crystallized using the hanging-drop method with 2-methylpentane-2,4-diol (MPD) as the precipitant. X-ray diffraction data have been collected on these crystals using a MAR Research imaging-plate detector system attached to a Rigaku RU200 rotating-anode X-ray generator. The crystals belong to the hexagonal space group P6122 with unit-cell dimensions of a = b = 96.21, c = 65.48 A. Preliminary molecular-replacement computations suggest that the structure of this protein is likely to be similar to that of calmodulin (CAM).

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Calcium-Binding Proteins / chemistry*
  • Calcium-Binding Proteins / isolation & purification
  • Crystallization
  • Crystallography, X-Ray
  • Drosophila melanogaster / chemistry
  • Entamoeba histolytica / chemistry*
  • Models, Molecular
  • Molecular Sequence Data
  • Molecular Weight
  • Protein Conformation*
  • Protozoan Proteins / chemistry*
  • Protozoan Proteins / isolation & purification
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Species Specificity

Substances

  • Calcium-Binding Proteins
  • Protozoan Proteins
  • Recombinant Fusion Proteins
  • calcium-binding protein, Entamoeba histolytica