Cloning, expression, purification and preliminary crystallographic data for Rv3214 (EntD), a predicted cofactor-dependent phosphoglycerate mutase from Mycobacterium tuberculosis

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Aug 1;61(Pt 8):753-5. doi: 10.1107/S1744309105020646. Epub 2005 Jul 8.

Abstract

The Mycobacterium tuberculosis open reading frame Rv3214, annotated as a cofactor-dependent phosphoglycerate mutase, has been cloned and expressed as an N-terminally His-tagged protein. Tagged, untagged and selenomethionine-labelled forms of Rv3214 (EntD) have been purified using nickel-affinity chromatography and gel filtration. The selenomethionine-labelled crystals diffracted to 2.15 A resolution and belong to space group P2(1), with unit-cell parameters a = 44.36, b = 79.03, c = 52.85 A, beta = 109.11 degrees. There are two molecules of molecular weight 21,948 Da per asymmetric unit.

MeSH terms

  • Cloning, Molecular
  • Coenzymes / metabolism*
  • Crystallography, X-Ray
  • Gene Expression*
  • Molecular Weight
  • Mycobacterium tuberculosis / enzymology*
  • Phosphoglycerate Mutase / chemistry*
  • Phosphoglycerate Mutase / genetics
  • Phosphoglycerate Mutase / isolation & purification*
  • Recombinant Fusion Proteins / chemistry
  • Scattering, Radiation

Substances

  • Coenzymes
  • Recombinant Fusion Proteins
  • Phosphoglycerate Mutase