Overexpression, crystallization and preliminary X-ray crystallographic analysis of a putative transposase from Thermoplasma acidophilum encoded by the Ta0474 gene

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Nov 1;62(Pt 11):1147-9. doi: 10.1107/S174430910604245X. Epub 2006 Oct 20.

Abstract

IS200 transposases, originally identified in Salmonella typhimurium LT2, are present in many bacteria and archaea and are distinct from other groups of transposases. To facilitate further structural comparisons among IS200-like transposases, structural analysis has been initiated of a putative transposase from Thermoplasma acidophilum encoded by the Ta0474 gene. Its 137-residue polypeptide shows high levels of sequence similarity to other members of the IS200 transposase family. The protein was overexpressed in intact form in Escherichia coli and crystallized at 297 K using a reservoir solution consisting of 100 mM Na HEPES pH 7.5 and 20%(v/v) ethanol. X-ray diffraction data were collected to 1.78 A. The crystals belong to the monoclinic space group P2(1), with unit-cell parameters a = 65.00, b = 34.07, c = 121.58 A, alpha = 90, beta = 100.20, gamma = 90 degrees. Four monomers, representing two copies of a dimeric molecule, are present in the asymmetric unit, giving a crystal volume per protein weight (V(M)) of 2.02 A(3) Da(-1) and a solvent content of 39.2%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaeal Proteins / chemistry
  • Base Sequence
  • Crystallography, X-Ray
  • DNA Primers
  • Genes, Archaeal*
  • Molecular Sequence Data
  • Thermoplasma / genetics*
  • Transposases / chemistry*
  • Transposases / genetics*
  • Transposases / isolation & purification
  • X-Ray Diffraction

Substances

  • Archaeal Proteins
  • DNA Primers
  • Transposases

Associated data

  • PDB/2A6M
  • PDB/2F4F