TY - JOUR
T1 - Structure and Mechanism of the Monoterpene Cyclolavandulyl Diphosphate Synthase that Catalyzes Consecutive Condensation and Cyclization
AU - Tomita, Takeo
AU - Kobayashi, Masaya
AU - Karita, Yuma
AU - Yasuno, Yoko
AU - Shinada, Tetsuro
AU - Nishiyama, Makoto
AU - Kuzuyama, Tomohisa
N1 - Funding Information:
We are grateful to Joseph P. Noel (The Salk Institute for Biological Studies) for advice on crystallography and to the staff of the Photon Factory for their assistance with data collection, which was approved by the Photon Factory Program Advisory Committee (2010G004, 2012G019, and 2014G106). We also thank Naoki Mori (The University of Tokyo) for the optical rotation of 6’. This work was supported by JSPS KAKENHI (26292058 and 16H06453 to T.K., and 17H05448 and 15K12758 to T.S.) and the JSPS A3 Foresight Program. M.K. was supported by a Grant-in-Aid for JSPS fellows (15J10131).
Publisher Copyright:
© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2017/11/20
Y1 - 2017/11/20
N2 - We report the three-dimensional structure of cyclolavandulyl diphosphate (CLPP) synthase (CLDS), which consecutively catalyzes the condensation of two molecules of dimethylallyl diphosphate (DMAPP) followed by cyclization to form a cyclic monoterpene, CLPP. The structures of apo-CLDS and CLDS in complex with Tris, pyrophosphate, and Mg2+ ion were refined at 2.00 Å resolution and 1.73 Å resolution, respectively. CLDS adopts a typical fold for cis-prenyl synthases and forms a homo-dimeric structure. An in vitro reaction using a regiospecifically 2H-substituted DMAPP substrate revealed the intramolecular proton transfer mechanism of the CLDS reaction. The CLDS structure and structure-based mutagenesis provide mechanistic insights into this unprecedented terpene synthase. The combination of structural and mechanistic insights advances the knowledge of intricate terpene synthase-catalyzed reactions.
AB - We report the three-dimensional structure of cyclolavandulyl diphosphate (CLPP) synthase (CLDS), which consecutively catalyzes the condensation of two molecules of dimethylallyl diphosphate (DMAPP) followed by cyclization to form a cyclic monoterpene, CLPP. The structures of apo-CLDS and CLDS in complex with Tris, pyrophosphate, and Mg2+ ion were refined at 2.00 Å resolution and 1.73 Å resolution, respectively. CLDS adopts a typical fold for cis-prenyl synthases and forms a homo-dimeric structure. An in vitro reaction using a regiospecifically 2H-substituted DMAPP substrate revealed the intramolecular proton transfer mechanism of the CLDS reaction. The CLDS structure and structure-based mutagenesis provide mechanistic insights into this unprecedented terpene synthase. The combination of structural and mechanistic insights advances the knowledge of intricate terpene synthase-catalyzed reactions.
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U2 - 10.1002/anie.201708474
DO - 10.1002/anie.201708474
M3 - Article
C2 - 28922556
AN - SCOPUS:85031091208
SN - 1433-7851
VL - 56
SP - 14913
EP - 14917
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 47
ER -