9M7E

Crystal structure of AsDMS D333N mutant in complex with drimenyl phosphate

これはPDB形式変換不可エントリーです。

9M7E の概要

• エントリーDOI: 10.2210/pdb9m7e/pdb
• 分子名称: Haloacid dehalogenase superfamily, subfamily IA, variant 3 with third motif having DD or ED, FARNESYL DIPHOSPHATE, CALCIUM ION, ... (7 entities in total)
• 機能のキーワード: terpene cyclase, haloacid dehalogenase, phosphatase, biosynthesis, biosynthetic protein
• 由来する生物種: Aquimarina spongiae
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 123148.71

構造登録者

Fujiyama, K.,Vo, N.N.Q.,Takahashi, S. (登録日: 2025-03-10, 公開日: 2025-09-10)

主引用文献

Fujiyama, K.,Takagi, H.,Vo, N.N.Q.,Morita, N.,Nogawa, T.,Takahashi, S.
Structural insights into a bacterial terpene cyclase fused with haloacid Dehalogenase-like phosphatase.
Chem Sci, 16:15310-15319, 2025

PubMed Abstract: Terpene cyclases (TCs), consisting of various combinations of α, β, and γ domains, have been extensively studied. Recently, non-canonical enzymes comprising a TCβ domain and a haloacid dehalogenase (HAD)-like domain (referred to as HAD-TCβ) have been discovered. However, their overall structure remains unclear. In this study, we determined the co-crystal structures of drimenol synthase from (AsDMS), which catalyzes the conversion of farnesyl pyrophosphate (1) into drimenol (2). Crystallographic analyses of the enzyme bound to substrates 1 and drimenyl monophosphate (3) demonstrated that the TCβ domain catalyzes a class II cyclization reaction initiated by protonation, whereas the HAD domain catalyzes a phosphatase-like dephosphorylation reaction dependent on a divalent metal. Crystallographic and gel filtration analyses revealed that AsDMS adopts a dimeric assembly. This dimerization positioned the TCβ and HAD domains to facilitate efficient substrate transfer electrostatic substrate channeling. Furthermore, to investigate the structure-function relationship of the AsDMS TCβ domain, we used AlphaFold2 to model the structure of the fungal albicanol (4) synthase. Comparative analysis of active-site residues between AsDMS and fungal 4-synthase enabled rational protein engineering, converting AsDMS activity from 2-synthase to 4-synthase. This study provides insights into the biosynthesis of valuable drimane-type sesquiterpenes targeted mutagenesis.

PubMed: 40852458
DOI: 10.1039/d5sc04719f

実験手法

X-RAY DIFFRACTION (2.9000384384 Å)