Crystallization and preliminary X-ray analysis of allene oxide synthase, cytochrome P450 CYP74A2, from Parthenium argentatum.
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.
Chang
Zhenzhan
Z
Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA.
Li
Lenong
L
Pan
Zhiqiang
Z
Wang
Xiaoqiang
X
D019659:Asteraceae; D003460:Crystallization / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000302:isolation & purification; D019746:Intramolecular Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D014961:X-Ray Diffraction
D010936:Plant Extracts; D010940:Plant Proteins; D003577:Cytochrome P-450 Enzyme System; D019746:Intramolecular Oxidoreductases; C021377:hydroperoxide isomerase