Crystallization and preliminary X-ray analysis of allene oxide synthase, cytochrome P450 CYP74A2, from Parthenium argentatum

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Jul 1;64(Pt 7):668-70. doi: 10.1107/S1744309108017545. Epub 2008 Jun 28.

Abstract

Oxylipins are oxygenated derivatives of fatty acids and pivotal signaling molecules in plants and animals. Allene oxide synthase (AOS) is a key cytochrome P450 CYP74 enzyme involved in the biosynthesis of plant oxylipin jasmonates to convert 13(S)-hydroperoxide to allene oxide. Guayule (Parthenium argentatum) AOS, CYP74A2, was expressed in Escherichia coli. Protein was purified using affinity chromatography and size exclusion chromatography, and then crystallized. Two different crystal forms were obtained from 0.2 M (NH(4))H(2)PO(4), 50% MPD, 0.1 M Tris, pH 8.5 at 277 K using the hanging-drop vapor-diffusion method. Preliminary X-ray analysis was carried out, and the crystals were found to belong to the tetragonal space group I422 with cell parameters a = b = 126.5, c = 163.9 A, and the monoclinic space group C2 with cell parameters a = 336.5, b = 184.2, c = 159.0 A, beta = 118.6 degrees . Diffraction data were collected to 2.4 A resolution from a tetragonal form of crystal using a home X-ray source.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Asteraceae*
  • Crystallization / methods
  • Cytochrome P-450 Enzyme System / chemistry*
  • Cytochrome P-450 Enzyme System / isolation & purification
  • Intramolecular Oxidoreductases / chemistry*
  • Intramolecular Oxidoreductases / isolation & purification
  • Plant Extracts / chemistry
  • Plant Extracts / isolation & purification
  • Plant Proteins / chemistry*
  • Plant Proteins / isolation & purification
  • X-Ray Diffraction*

Substances

  • Plant Extracts
  • Plant Proteins
  • Cytochrome P-450 Enzyme System
  • Intramolecular Oxidoreductases
  • hydroperoxide isomerase