5YQT

Crystal Structure of the L74F/M78V/I80V/L114F mutant of LEH complexed with cyclopentene oxide

5YQT の概要

• エントリーDOI: 10.2210/pdb5yqt/pdb
• 分子名称: Limonene-1,2-epoxide hydrolase, (1R,5S)-6-oxabicyclo[3.1.0]hexane (3 entities in total)
• 機能のキーワード: epoxide hydrolase, hydrolase
• 由来する生物種: Rhodococcus erythropolis (Arthrobacter picolinophilus)
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 139375.63

構造登録者

Kong, X.D.,Sun, Z.T.,Wu, L.,Reetz, M.T.,Zhou, J.H.,Xu, J.H. (登録日: 2017-11-07, 公開日: 2018-06-27, 最終更新日: 2023-11-22)

主引用文献

Sun, Z.,Wu, L.,Bocola, M.,Chan, H.C.S.,Lonsdale, R.,Kong, X.D.,Yuan, S.,Zhou, J.,Reetz, M.T.
Structural and Computational Insight into the Catalytic Mechanism of Limonene Epoxide Hydrolase Mutants in Stereoselective Transformations.
J. Am. Chem. Soc., 140:310-318, 2018

PubMed Abstract: Directed evolution of limonene epoxide hydrolase (LEH), which catalyzes the hydrolytic desymmetrization reactions of cyclopentene oxide and cyclohexene oxide, results in (R,R)- and (S,S)-selective mutants. Their crystal structures combined with extensive theoretical computations shed light on the mechanistic intricacies of this widely used enzyme. From the computed activation energies of various pathways, we discover the underlying stereochemistry for favorable reactions. Surprisingly, some of the most enantioselective mutants that rapidly convert cyclohexene oxide do not catalyze the analogous transformation of the structurally similar cyclopentene oxide, as shown by additional X-ray structures of the variants harboring this slightly smaller substrate. We explain this puzzling observation on the basis of computational calculations which reveal a disrupted alignment between nucleophilic water and cyclopentene oxide due to the pronounced flexibility of the binding pocket. In contrast, in the stereoselective reactions of cyclohexene oxide, reactive conformations are easily reached. The unique combination of structural and computational data allows insight into mechanistic details of this epoxide hydrolase and provides guidance for future protein engineering in reactions of structurally different substrates.

PubMed: 29232125
DOI: 10.1021/jacs.7b10278

実験手法

X-RAY DIFFRACTION (2.3 Å)