8E9C

Crystal structure of E. coli aspartate aminotransferase mutant AIFS in the ligand-free form at 100 K

8E9C の概要

• エントリーDOI: 10.2210/pdb8e9c/pdb
• 分子名称: Aspartate aminotransferase, PYRIDOXAL-5'-PHOSPHATE, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: enzyme, designed protein, mutant protein, transferase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 90189.25

構造登録者

Chica, R.A.,St-Jacques, A.D.,Rodriguez, J.M.,Thompson, M.C. (登録日: 2022-08-26, 公開日: 2022-11-02, 最終更新日: 2023-10-18)

主引用文献

St-Jacques, A.D.,Rodriguez, J.M.,Eason, M.G.,Foster, S.M.,Khan, S.T.,Damry, A.M.,Goto, N.K.,Thompson, M.C.,Chica, R.A.
Computational remodeling of an enzyme conformational landscape for altered substrate selectivity.
Nat Commun, 14:6058-6058, 2023

PubMed Abstract: Structural plasticity of enzymes dictates their function. Yet, our ability to rationally remodel enzyme conformational landscapes to tailor catalytic properties remains limited. Here, we report a computational procedure for tuning conformational landscapes that is based on multistate design of hinge-mediated domain motions. Using this method, we redesign the conformational landscape of a natural aminotransferase to preferentially stabilize a less populated but reactive conformation and thereby increase catalytic efficiency with a non-native substrate, resulting in altered substrate selectivity. Steady-state kinetics of designed variants reveals activity increases with the non-native substrate of approximately 100-fold and selectivity switches of up to 1900-fold. Structural analyses by room-temperature X-ray crystallography and multitemperature nuclear magnetic resonance spectroscopy confirm that conformational equilibria favor the target conformation. Our computational approach opens the door to targeted alterations of conformational states and equilibria, which should facilitate the design of biocatalysts with customized activity and selectivity.

PubMed: 37770431
DOI: 10.1038/s41467-023-41762-0

実験手法

X-RAY DIFFRACTION (2.18 Å)