Crystallization and characterization of a fragment of pseudouridine synthase RluC from Escherichia coli

Acta Crystallogr D Biol Crystallogr. 1999 Jan;55(Pt 1):302-4. doi: 10.1107/S090744499801021X. Epub 1999 Jan 1.

Abstract

RluC from E. coli is the enzyme responsible for catalyzing the isomerization of uridines 955, 2504 and 2580 in 23S rRNA to pseudouridine. Histidine-tagged RluC was cloned, overexpressed and purified by nickel-affinity chromatography. A proteolytically derived fragment of the enzyme consisting of residues 89-319 has been shown to retain catalytic activity. Crystals of this fragment, grown by precipitation with sodium acetate at pH 8.0, belong to space group P321, with unit-cell dimensions a = b = 97.1, c = 86.3 A and have two molecules in the crystallographic asymmetric unit. The flash-frozen crystals diffract X-rays to at least 2.3 A resolution and appear suitable for crystal structure determination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Gene Expression
  • Hydro-Lyases / chemistry*
  • Hydro-Lyases / genetics
  • Hydro-Lyases / isolation & purification*
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / isolation & purification
  • RNA, Bacterial
  • RNA, Ribosomal, 23S
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification

Substances

  • Peptide Fragments
  • RNA, Bacterial
  • RNA, Ribosomal, 23S
  • Recombinant Proteins
  • Hydro-Lyases
  • pseudouridylate synthetase