Purification, crystallization and preliminary X-ray diffraction data from selenomethionine glycinamide ribonucleotide synthetase

Acta Crystallogr D Biol Crystallogr. 1999 Feb;55(Pt 2):518-21. doi: 10.1107/s0907444998009020.

Abstract

In this study, the overexpression, purification and crystallization of selenomethionine (SeMet) incorporated glycinamide ribonucleotide synthetase (GAR-syn) from Escherichia coli are reported. The overexpression of SeMet GAR-syn was placed under the control of the isopropylthio-beta-galactoside (IPTG) inducible T7 RNA-polymerase system. The newly developed construct contained a removable histidine tag on the amino terminus of GAR-syn, which allowed rapid purification using metal-chelate chromatography techniques. The SeMet GAR-syn crystals were grown by hanging-drop vapor diffusion and belong to the space group P212121 with unit-cell parameters a = 56.2, b = 62.4 and c = 129.8 A and a single monomer in the asymmetric unit. The crystals diffract to 1.6 A resolution and have led to the determination of multiple-wavelength anomalous diffraction phases to 2.2 A resolution.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Carbon-Nitrogen Ligases / chemistry*
  • Carbon-Nitrogen Ligases / isolation & purification
  • Crystallization
  • Crystallography, X-Ray
  • DNA Primers
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Selenium / chemistry

Substances

  • DNA Primers
  • Recombinant Proteins
  • Carbon-Nitrogen Ligases
  • selenomethionine glycinamide ribonucleotide synthetase
  • Selenium