Purification, crystallization and preliminary X-ray analysis of glucokinase from Streptomyces griseus in complex with glucose

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Aug 1;67(Pt 8):914-6. doi: 10.1107/S1744309111022275. Epub 2011 Jul 20.

Abstract

Glucokinase catalyzes the phosphorylation of glucose using ATP to yield glucose 6-phosphate. SgGlkA is a bacterial group III glucokinase from Streptomyces griseus that seems to play a regulatory role in carbon catabolite repression in this organism. SgGlkA was expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method at 293 K. A crystal of SgGlkA in complex with glucose was obtained using a reservoir solution consisting of 0.9 M sodium/potassium tartrate, 0.2 M NaCl and 0.1 M imidazole pH 8.1 and diffracted X-rays to 1.84 Å resolution. The crystal of SgGlkA in complex with glucose belonged to space group P6(2)22 or P6(4)22, with unit-cell parameters a = b = 109.19, c = 141.18 Å. The crystal contained one molecule in the asymmetric unit.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization
  • Crystallography, X-Ray
  • Glucokinase / chemistry*
  • Glucokinase / isolation & purification
  • Glucokinase / metabolism
  • Glucose / chemistry*
  • Glucose / metabolism
  • Protein Binding
  • Streptomyces griseus / enzymology*

Substances

  • Glucokinase
  • Glucose