Expression, purification, crystallization and preliminary X-ray analysis of glucose-1-phosphate uridylyltransferase (GalU) from Erwinia amylovora

Acta Crystallogr F Struct Biol Commun. 2014 Sep;70(Pt 9):1249-51. doi: 10.1107/S2053230X14016458. Epub 2014 Aug 27.

Abstract

Glucose-1-phosphate uridylyltransferase from Erwinia amylovora CFPB1430 was expressed as a His-tag fusion protein in Escherichia coli. After tag removal, the purified protein was crystallized from 100 mM Tris pH 8.5, 2 M ammonium sulfate, 5% ethylene glycol. Diffraction data sets were collected to a maximum resolution of 2.46 Å using synchrotron radiation. The crystals belonged to the hexagonal space group P62, with unit-cell parameters a = 80.67, b = 80.67, c = 169.18. The structure was solved by molecular replacement using the structure of the E. coli enzyme as a search model.

Keywords: Erwinia; GalU; UDP-glucose; amylovoran; sugar metabolism; uridylyltransferase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • Crystallography, X-Ray / methods*
  • DNA Primers
  • Erwinia amylovora / enzymology*
  • Polymerase Chain Reaction
  • Protein Conformation
  • UTP-Glucose-1-Phosphate Uridylyltransferase / chemistry*
  • UTP-Glucose-1-Phosphate Uridylyltransferase / genetics
  • UTP-Glucose-1-Phosphate Uridylyltransferase / isolation & purification

Substances

  • DNA Primers
  • UTP-Glucose-1-Phosphate Uridylyltransferase