7WCC

Oxidase ChaP-D49L/Q91C mutant

7WCC の概要

• エントリーDOI: 10.2210/pdb7wcc/pdb
• 分子名称: ChaP, FE (III) ION (3 entities in total)
• 機能のキーワード: chap, oxidative rearrangement steps, mycobacterium tuberculosis, biosynthetic protein
• 由来する生物種: Streptomyces chartreusis
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 57888.37

構造登録者

Sun, M.X.,Zheng, W.,Wang, Y.S.,Zhu, J.P.,Tan, R.X. (登録日: 2021-12-19, 公開日: 2023-05-24, 最終更新日: 2024-12-18)

主引用文献

Wang, Y.S.,Zheng, W.,Jiang, N.,Jin, Y.X.,Meng, Z.K.,Sun, M.X.,Zong, Y.L.,Xu, T.,Zhu, J.,Tan, R.X.
Alteration of the Catalytic Reaction Trajectory of a Vicinal Oxygen Chelate Enzyme by Directed Evolution.
Angew.Chem.Int.Ed.Engl., 61:e202201321-e202201321, 2022

PubMed Abstract: The vicinal oxygen chelate (VOC) metalloenzyme superfamily catalyzes a highly diverse set of reactions with the mechanism characterized by the bidentate coordination of vicinal oxygen atoms to metal ion centers, but there remains a lack of a platform to steer the reaction trajectories, especially for o-quinone metabolizing pathways. Herein, we present the directed-evolution-enabled bifunctional turnover of ChaP, which is a homotetramer and represents an unprecedented VOC enzyme class. Unlike the ChaP catalysis of extradiol-like o-quinone cleavage and concomitant α-keto acid decarboxylation, a group of ChaP variants (CVs) catalyze intradiol-like o-quinone deconstruction and CO liberation from the resulting o-hydroxybenzoic acid scaffolds with high regioselectivity. Enzyme crystal structures, labeling experiments and computational simulations corroborated that the D49L mutation allows the metal ion to change its coordination with the tyrosine phenoxy atoms in different monomers, thereby altering the reaction trajectory with the regiospecificity further improved by the follow-up replacement of the Y92 residue with any of alanine, glycine, threonine, and serine. The study highlights the unpredicted catalytic versatility and enzymatic plasticity of VOC enzymes with biotechnological significance.

PubMed: 35415958
DOI: 10.1002/anie.202201321

実験手法

X-RAY DIFFRACTION (1.5 Å)