Improved expression, purification and crystallization of a putative N-acetyl-gamma-glutamyl-phosphate reductase from rice (Oryza sativa)

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Dec 1;61(Pt 12):1058-61. doi: 10.1107/S1744309105035384. Epub 2005 Nov 24.

Abstract

N-Acetyl-gamma-glutamyl-phosphate reductase (AGPR) catalyzes the third step in an eight-step arginine-biosynthetic pathway that starts with glutamate. This enzyme converts N-acetyl-gamma-glutamyl phosphate to N-acetylglutamate-gamma-semialdehyde by an NADPH-dependent reductive dephosphorylation. AGPR from Oryza sativa (OsAGPR) was expressed in Escherichia coli at 291 K as a soluble fusion protein with an upstream thioredoxin-hexahistidine [Trx-(His)6] extension. OsAGPR(Ala50-Pro366) was purified and crystals were obtained using the sitting-drop vapour-diffusion method at 293 K and diffract X-rays to at least 1.8 A resolution. They belong to the hexagonal space group P6(1), with unit-cell parameters a = 86.11, c = 316.3 A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Oxidoreductases / chemistry*
  • Amino Acids / chemistry
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Diffusion
  • Escherichia coli / metabolism
  • Glutamates / chemistry
  • Histidine / chemistry
  • Humans
  • NADP / chemistry
  • Oligopeptides / chemistry
  • Oryza / enzymology*
  • Protein Conformation
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Proteins / chemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Temperature
  • Thioredoxins / chemistry
  • X-Ray Diffraction

Substances

  • Amino Acids
  • Glutamates
  • His-His-His-His-His-His
  • Oligopeptides
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Histidine
  • Thioredoxins
  • NADP
  • Aldehyde Oxidoreductases
  • N-acetyl-gamma-glutamyl-phosphate reductase