Crystallization and preliminary X-ray crystallographic analysis of Escherichia coli CyaY, a structural homologue of human frataxin

Acta Crystallogr D Biol Crystallogr. 2000 Jul;56(Pt 7):920-1. doi: 10.1107/s0907444900005916.

Abstract

CyaY is a 106-residue protein from Escherichia coli. It shows amino-acid sequence similarity to human frataxin and a frataxin homologue in Saccharomyces cerevisiae, Yfh1p. The former is associated with the disease Friedreich ataxia and the latter plays a key role in iron homeostasis in mitochondria. CyaY has been overexpressed in soluble form in E. coli. The recombinant protein with a His(6) tag at its C-terminus has been crystallized at 296 K using polyethylene glycol (PEG) 4000 as a precipitant. Native diffraction data have been collected to 1.8 A using Cu Kalpha X-rays. The crystals belong to the trigonal space group P3(1)21 (or P3(2)21), with unit-cell parameters a = b = 44.66, c = 99.87 A, alpha = beta = 90.0, gamma = 120.0 degrees. The asymmetric unit contains one molecule of recombinant CyaY, with a corresponding V(m) of 2.13 A(3) Da(-1) and solvent content of 42.3%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / chemistry*
  • Escherichia coli Proteins
  • Frataxin
  • Humans
  • Iron-Binding Proteins*
  • Phosphotransferases (Alcohol Group Acceptor) / chemistry*
  • Protein Conformation
  • Recombinant Proteins / chemistry

Substances

  • Bacterial Proteins
  • CyaY protein, E coli
  • Escherichia coli Proteins
  • Iron-Binding Proteins
  • Recombinant Proteins
  • Phosphotransferases (Alcohol Group Acceptor)