Purification, crystallization and preliminary X-ray diffraction analysis of pathogen-inducible oxygenase (PIOX) from Oryza sativa

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Apr 1;62(Pt 4):365-7. doi: 10.1107/S1744309106007305. Epub 2006 Mar 10.

Abstract

Pathogen-inducible oxygenase (PIOX) is a heme-containing membrane-associated protein found in monocotyledon and dicotyledon plants that utilizes molecular oxygen to convert polyunsaturated fatty acids into their corresponding 2R-hydroperoxides. PIOX is a member of a larger family of fatty-acid alpha-dioxygenases that includes the mammalian cyclooxygenase enzymes cyclooxygenase 1 and 2 (COX-1 and COX-2). Single crystals of PIOX from rice (Oryza sativa) have been grown from MPD using recombinant protein expressed in Escherichia coli and subsequently extracted utilizing decyl maltoside as the solubilizing detergent. Crystals diffract to 3.0 angstroms resolution using a rotating-anode generator and R-AXIS IV detector, and belong to space group P1. Based on the Matthews coefficient and self-rotation function analyses, there are presumed to be four molecules in the asymmetric unit related by noncrystallographic 222 symmetry.

Publication types

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

MeSH terms

  • Crystallization
  • Dioxygenases / chemistry*
  • Dioxygenases / isolation & purification
  • Dioxygenases / metabolism
  • Electron Transport Complex IV / chemistry
  • Electron Transport Complex IV / isolation & purification
  • Electron Transport Complex IV / metabolism
  • Linoleic Acid / metabolism
  • Macromolecular Substances
  • Membrane Proteins / chemistry*
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism
  • Oryza / enzymology*
  • Plant Proteins / chemistry*
  • Plant Proteins / isolation & purification
  • Plant Proteins / metabolism
  • X-Ray Diffraction

Substances

  • Macromolecular Substances
  • Membrane Proteins
  • Plant Proteins
  • Linoleic Acid
  • Dioxygenases
  • Electron Transport Complex IV