Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the small subunit of isopropylmalate isomerase (Rv2987c) from Mycobacterium tuberculosis

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Feb 1;65(Pt 2):136-9. doi: 10.1107/S1744309108042516. Epub 2009 Jan 31.

Abstract

Two C-terminally truncated variants of the small subunit of Mycobacterium tuberculosis isopropylmalate isomerase (Rv2987c; LeuD), LeuD_1-156 and LeuD_1-168, have been cloned, heterologously expressed in Escherichia coli, purified using standard chromatographic techniques and crystallized. The crystals of LeuD_1-156 belonged to the hexagonal system (space group P6(1)22 or P6(5)22) with up to four subunits in the asymmetric unit, whereas the crystals of LeuD_1-168 belonged to the monoclinic system (space group P2(1)) with two subunits in the asymmetric unit. Both crystals diffracted X-rays to beyond 2.0 A resolution and were suitable for further crystallographic analysis.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Crystallization
  • Gene Expression Regulation, Bacterial / physiology
  • Isomerases / biosynthesis
  • Isomerases / genetics
  • Isomerases / isolation & purification*
  • Mycobacterium tuberculosis / enzymology*
  • Mycobacterium tuberculosis / genetics*
  • Peptide Fragments / biosynthesis
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / isolation & purification
  • Protein Folding
  • Ribosome Subunits, Small, Bacterial / enzymology*
  • Ribosome Subunits, Small, Bacterial / genetics*
  • X-Ray Diffraction* / methods

Substances

  • Peptide Fragments
  • isopropylmalate isomerase
  • Isomerases