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. 2005 Aug 1;61(Pt 8):799-801.
doi: 10.1107/S1744309105023365. Epub 2005 Jul 30.

Crystallization of Delta1-tetrahydrocannabinolic acid (THCA) synthase from Cannabis sativa

Yoshinari Shoyama  1 , Ayako Takeuchi, Futoshi Taura, Taro Tamada, Motoyasu Adachi, Ryota Kuroki, Yukihiro Shoyama, Satoshi Morimoto
Affiliations

Crystallization of Delta1-tetrahydrocannabinolic acid (THCA) synthase from Cannabis sativa

Yoshinari Shoyama et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. .

Abstract

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.

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Figures

Figure 1
Figure 1
THCA biosynthesis. THCA synthase catalyzes oxidative cyclization of the monoterpene moiety of CBGA to form THCA. THC is derived from THCA by non-enzymatic decarboxylation.
Figure 2
Figure 2
Crystals of THCA synthase. The scale bar is 0.1 mm in length.
See this image and copyright information in PMC

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