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Title: Crystallization and Preliminary X-ray Analysis of Allene Oxide Synthase, Cytochrome P450 CYP74A2, from Parthenium argentatum

Author
item CHANG, ZHENZHAN - SAMUEL ROBERTS NOBLE FOUN
item LI, LENONG - SAMUEL ROBERTS NOBLE FOUN
item Pan, Zhiqiang - Peter
item WANG, XIAOQIANG - SAMUEL ROBERTS NOBLE FOUN

Submitted to: Acta Crystallographica Section F: Structural Biology Communications
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/10/2008
Publication Date: 6/28/2008
Citation: Chang, Z., Li, L., Pan, Z., Wang, X. 2008. Crystallization and Preliminary X-ray Analysis of Allene Oxide Synthase, Cytochrome P450 CYP74A2, from Parthenium argentatum. Acta Crystallographica Section F. F64:668-670.

Interpretive Summary: 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 jasmonates in oxylipin biosynthetic pathway, in which 13(S)-hydroperoxide is converted to allene oxide which is further cyclized by allene oxide cyclase, leading to the formation of jasmonic acid. In order to study the enzyme-substrate interaction, the AOS enzyme from guayule (Parthenium argentatum) was expressed in E. coli., purified and crystallized. This paper reported the preliminary X-ray analysis and diffraction data collected to 2.4 Å resolution from a tetragonal form of crystal using a home X-ray source.

Technical 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 E. coli. Protein was purified using affinity chromatography and size exclusion chromatography, and then crystallized. Two different crystal forms were obtained from 0.2 M (NH4)H2PO4, 50% MPD, 0.1 M Tris, pH8.5 at 277 K using the hanging drop vapor-diffusion method. Preliminary X-ray analysis was carried out, and the crystals belong to the tetragonal space group I422 with cell parameters a=b=126.5, c=163.9Å, and the monoclinic space group C2 with cell parameters a=336.5, b=184.2, c=159.0Å, B=118.6, respectively. Diffraction data were collected to 2.4 Å resolution from a tetragonal form of crystal using a home X-ray source.