7OLP

Dioxygenase AsqJ mutant (V72I) in complex with 2 and alpha-ketoglutarate

7OLP の概要

• エントリーDOI: 10.2210/pdb7olp/pdb
• 関連するPDBエントリー: 5DAP
• 分子名称: Iron/alpha-ketoglutarate-dependent dioxygenase asqJ, NICKEL (II) ION, 2-OXOGLUTARIC ACID, ... (5 entities in total)
• 機能のキーワード: quinolone biosynthesis, molecular engineering, epoxidation, catalytic mechanism, oxidoreductase
• 由来する生物種: Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139) (Aspergillus nidulans)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34577.09

構造登録者

Auman, D.,Mader, S.L.,Ecker, F.,Dorst, K.,Braeuer, A.,Widmalm, G.,Groll, M.,Kaila, V.R.I. (登録日: 2021-05-20, 公開日: 2022-06-01, 最終更新日: 2024-01-31)

主引用文献

Auman, D.,Ecker, F.,Mader, S.L.,Dorst, K.M.,Brauer, A.,Widmalm, G.,Groll, M.,Kaila, V.R.I.
Peroxy Intermediate Drives Carbon Bond Activation in the Dioxygenase AsqJ.
J.Am.Chem.Soc., 144:15622-15632, 2022

PubMed Abstract: Dioxygenases catalyze stereoselective oxygen atom transfer in metabolic pathways of biological, industrial, and pharmaceutical importance, but their precise chemical principles remain controversial. The α-ketoglutarate (αKG)-dependent dioxygenase AsqJ synthesizes biomedically active quinolone alkaloids via desaturation and subsequent epoxidation of a carbon-carbon bond in the cyclopeptin substrate. Here, we combine high-resolution X-ray crystallography with enzyme engineering, quantum-classical (QM/MM) simulations, and biochemical assays to describe a peroxidic intermediate that bridges the substrate and active site metal ion in AsqJ. Homolytic cleavage of this moiety during substrate epoxidation generates an activated high-valent ferryl (Fe = O) species that mediates the next catalytic cycle, possibly without the consumption of the metabolically valuable αKG cosubstrate. Our combined findings provide an important understanding of chemical bond activation principles in complex enzymatic reaction networks and molecular mechanisms of dioxygenases.

PubMed: 35980821
DOI: 10.1021/jacs.2c05650

実験手法

X-RAY DIFFRACTION (1.55 Å)