5GUE

Crystal structure of CotB2 (GGSPP/Mg2+-Bound Form) from Streptomyces melanosporofaciens

5GUE の概要

• エントリーDOI: 10.2210/pdb5gue/pdb
• 関連するPDBエントリー: 5GUC
• 分子名称: Cyclooctat-9-en-7-ol synthase, phosphonooxy-[(10E)-3,7,11,15-tetramethylhexadeca-2,6,10,14-tetraenyl]sulfanyl-phosphinic acid, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: terpene cyclase fold, diterpene cyclase, lyase, biosynthetic protein
• 由来する生物種: Streptomyces melanosporofaciens
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 154731.77

構造登録者

Tomita, T.,Kim, S.-Y.,Ozaki, T.,Yoshida, A.,Kuzuyama, T.,Nishiyama, M. (登録日: 2016-08-28, 公開日: 2017-06-07, 最終更新日: 2023-11-08)

主引用文献

Tomita, T.,Kim, S.-Y.,Teramoto, K.,Meguro, A.,Ozaki, T.,Yoshida, A.,Motoyoshi, Y.,Mori, N.,Ishigami, K.,Watanabe, H.,Nishiyama, M.,Kuzuyama, T.
Structural Insights into the CotB2-Catalyzed Cyclization of Geranylgeranyl Diphosphate to the Diterpene Cyclooctat-9-en-7-ol
ACS Chem. Biol., 12:1621-1628, 2017

PubMed Abstract: The diterpene cyclase CotB2 catalyzes the cyclization of geranylgeranyl diphosphate (GGPP) to the tricyclic cyclooctat-9-en-7-ol, which is characterized by a 5-8-5-fused ring skeleton. We have previously proposed a cyclization cascade involving a unique carbon-carbon bond rearrangement combined with multiple hydride shifts, all occurring at a single active site. Here, we report the first high-resolution X-ray crystal structure of CotB2 with bound substrate analog geranylgeranyl thiodiphosphate (GGSPP). In the GGSPP-bound form, GGSPP folds into a unique S-shaped conformation that probably reflects the substrate-bound state prior to ionization of the substrate GGPP. The folded framework of GGSPP is surrounded by hydrophobic residues and several aromatic and asparagine residues that are well-positioned to stabilize a series of reactive carbocation intermediates through a combination of cation-π and dipole charge interactions. The combined crystal structures and mutagenesis-based biochemical assays provide a structural basis for exquisite control of ring formation and stereochemistry during CotB2 catalysis.

PubMed: 28463490
DOI: 10.1021/acschembio.7b00154

実験手法

X-RAY DIFFRACTION (1.8 Å)