2PS6

N225D/S229T trichodiene synthase

2PS6 の概要

• エントリーDOI: 10.2210/pdb2ps6/pdb
• 関連するPDBエントリー: 1JFA 1KIY 1YJ4 2AEK 2PS4 2PS7
• 分子名称: Trichodiene synthase, 1,2-ETHANEDIOL (3 entities in total)
• 機能のキーワード: terpenoid synthase fold, site-directed mutagenesis, nse/dte motif, magnesium, ethylene glycol, lyase
• 由来する生物種: Fusarium sporotrichioides
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 88255.36

構造登録者

Vedula, L.S.,Cane, D.E.,Christianson, D.W. (登録日: 2007-05-04, 公開日: 2007-12-18, 最終更新日: 2023-08-30)

主引用文献

Vedula, L.S.,Jiang, J.,Zakharian, T.,Cane, D.E.,Christianson, D.W.
Structural and mechanistic analysis of trichodiene synthase using site-directed mutagenesis: probing the catalytic function of tyrosine-295 and the asparagine-225/serine-229/glutamate-233-Mg2+B motif.
Arch.Biochem.Biophys., 469:184-194, 2008

PubMed Abstract: Trichodiene synthase from Fusarium sporotrichioides contains two metal ion-binding motifs required for the cyclization of farnesyl diphosphate: the "aspartate-rich" motif D(100)DXX(D/E) that coordinates to Mg2+A and Mg2+C, and the "NSE/DTE" motif N(225)DXXSXXXE that chelates Mg2+B (boldface indicates metal ion ligands). Here, we report steady-state kinetic parameters, product array analyses, and X-ray crystal structures of trichodiene synthase mutants in which the fungal NSE motif is progressively converted into a plant-like DDXXTXXXE motif, resulting in a degradation in both steady-state kinetic parameters and product specificity. Each catalytically active mutant generates a different distribution of sesquiterpene products, and three newly detected sesquiterpenes are identified. In addition, the kinetic and structural properties of the Y295F mutant of trichodiene synthase were found to be similar to those of the wild-type enzyme, thereby ruling out a proposed role for Y295 in catalysis.

PubMed: 17996718
DOI: 10.1016/j.abb.2007.10.015

実験手法

X-RAY DIFFRACTION (2.6 Å)