Crystallization of Delta1-tetrahydrocannabinolic acid (THCA) synthase from Cannabis sativa.
Delta1-Tetrahydrocannabinolic acid (THCA) synthase is a novel oxidoreductase that catalyzes the biosynthesis of the psychoactive compound THCA in Cannabis sativa (Mexican strain). In order to investigate the structure-function relationship of THCA synthase, this enzyme was overproduced in insect cells, purified and finally crystallized in 0.1 M HEPES buffer pH 7.5 containing 1.4 M sodium citrate. A single crystal suitable for X-ray diffraction measurement was obtained in 0.09 M HEPES buffer pH 7.5 containing 1.26 M sodium citrate. The crystal diffracted to 2.7 A resolution at beamline BL41XU, SPring-8. The crystal belonged to the primitive cubic space group P432, with unit-cell parameters a = b = c = 178.2 A. The calculated Matthews coefficient was approximately 4.1 or 2.0 A3 Da(-1) assuming the presence of one or two molecules of THCA synthase in the asymmetric unit, respectively.
Shoyama
Yoshinari
Y
Faculty of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582, Japan.
Takeuchi
Ayako
A
Taura
Futoshi
F
Tamada
Taro
T
Adachi
Motoyasu
M
Kuroki
Ryota
R
Shoyama
Yukihiro
Y
Morimoto
Satoshi
S
D002188:Cannabis / Q000201:enzymology*; D003460:Crystallization; D013759:Dronabinol / Q000096:biosynthesis / Q000737:chemistry; D019746:Intramolecular Oxidoreductases / Q000737:chemistry*
D013759:Dronabinol; D019746:Intramolecular Oxidoreductases; C100677:delta(1)-tetrahydrocannabinolic acid synthase