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

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 62; no. 4; pp. 365 - 367
• Main Authors: Lloyd, Tracy, Krol, Adam, Campanaro, Danielle, Malkowski, Michael
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Blackwell Publishing Ltd 01-04-2006; International Union of Crystallography
• Subjects: ANODES; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; Crystallization Communications; detergents; Dioxygenases - chemistry; Dioxygenases - isolation & purification; Dioxygenases - metabolism; Electron Transport Complex IV - chemistry; Electron Transport Complex IV - isolation & purification; Electron Transport Complex IV - metabolism; ESCHERICHIA COLI; fatty-acid α-dioxygenases; HEME; Linoleic Acid - metabolism; Macromolecular Substances; MEMBRANE PROTEINS; Membrane Proteins - chemistry; Membrane Proteins - isolation & purification; Membrane Proteins - metabolism; MEMBRANES; MOLECULES; MONOCRYSTALS; Oryza - enzymology; Oryza sativa; OXYGEN; PIOX; Plant Proteins - chemistry; Plant Proteins - isolation & purification; Plant Proteins - metabolism; RESOLUTION; ROTATION; SPACE GROUPS; SYMMETRY; X-RAY DIFFRACTION
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309106007305

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 α‐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 Å 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.