7K4X

Crystal structure of Kemp Eliminase HG3.7 in complex with the transition state analog 6-nitrobenzotriazole

7K4X の概要

• エントリーDOI: 10.2210/pdb7k4x/pdb
• 分子名称: Endo-1,4-beta-xylanase, 6-NITROBENZOTRIAZOLE, ACETATE ION, ... (5 entities in total)
• 機能のキーワード: kemp elimination, directed evolution, transition state analog, hydrolase
• 由来する生物種: Thermoascus aurantiacus
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 67759.76

構造登録者

Padua, R.A.P.,Otten, R.,Bunzel, A.,Nguyen, V.,Pitsawong, W.,Patterson, M.,Sui, S.,Perry, S.L.,Cohen, A.E.,Hilvert, D.,Kern, D. (登録日: 2020-09-16, 公開日: 2020-12-02, 最終更新日: 2023-10-18)

主引用文献

Otten, R.,Padua, R.A.P.,Bunzel, H.A.,Nguyen, V.,Pitsawong, W.,Patterson, M.,Sui, S.,Perry, S.L.,Cohen, A.E.,Hilvert, D.,Kern, D.
How directed evolution reshapes the energy landscape in an enzyme to boost catalysis.
Science, 370:1442-1446, 2020

PubMed Abstract: The advent of biocatalysts designed computationally and optimized by laboratory evolution provides an opportunity to explore molecular strategies for augmenting catalytic function. Applying a suite of nuclear magnetic resonance, crystallography, and stopped-flow techniques to an enzyme designed for an elementary proton transfer reaction, we show how directed evolution gradually altered the conformational ensemble of the protein scaffold to populate a narrow, highly active conformational ensemble and accelerate this transformation by nearly nine orders of magnitude. Mutations acquired during optimization enabled global conformational changes, including high-energy backbone rearrangements, that cooperatively organized the catalytic base and oxyanion stabilizer, thus perfecting transition-state stabilization. The development of protein catalysts for many chemical transformations could be facilitated by explicitly sampling conformational substates during design and specifically stabilizing productive substates over all unproductive conformations.

PubMed: 33214289
DOI: 10.1126/science.abd3623

実験手法

X-RAY DIFFRACTION (1.6 Å)