5KJC

V222I horse liver alcohol dehydrogenase complexed with NAD+ and pentafluorobenzyl alcohol

5KJC の概要

• エントリーDOI: 10.2210/pdb5kjc/pdb
• 関連するPDBエントリー: 4DWV 5KCP 5KCZ 5KJ1 5KJ6 5KJE 5KJF
• 分子名称: Alcohol dehydrogenase E chain, ZINC ION, NICOTINAMIDE-ADENINE-DINUCLEOTIDE (ACIDIC FORM), ... (6 entities in total)
• 機能のキーワード: oxidoreductase, rossmann fold, michaelis analogue, dynamics
• 由来する生物種: Equus caballus (Horse)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 82310.14

構造登録者

Plapp, B.V. (登録日: 2016-06-18, 公開日: 2016-06-29, 最終更新日: 2023-09-27)

主引用文献

Shanmuganatham, K.K.,Wallace, R.S.,Ting-I Lee, A.,Plapp, B.V.
Contribution of buried distal amino acid residues in horse liver alcohol dehydrogenase to structure and catalysis.
Protein Sci., 27:750-768, 2018

PubMed Abstract: The dynamics of enzyme catalysis range from the slow time scale (∼ms) for substrate binding and conformational changes to the fast time (∼ps) scale for reorganization of substrates in the chemical step. The contribution of global dynamics to catalysis by alcohol dehydrogenase was tested by substituting five different, conserved amino acid residues that are distal from the active site and located in the hinge region for the conformational change or in hydrophobic clusters. X-ray crystallography shows that the structures for the G173A, V197I, I220 (V, L, or F), V222I, and F322L enzymes complexed with NAD and an analogue of benzyl alcohol are almost identical, except for small perturbations at the sites of substitution. The enzymes have very similar kinetic constants for the oxidation of benzyl alcohol and reduction of benzaldehyde as compared to the wild-type enzyme, and the rates of conformational changes are not altered. Less conservative substitutions of these amino acid residues, such as G173(V, E, K, or R), V197(G, S, or T), I220(G, S, T, or N), and V222(G, S, or T) produced unstable or poorly expressed proteins, indicating that the residues are critical for global stability. The enzyme scaffold accommodates conservative substitutions of distal residues, and there is no evidence that fast, global dynamics significantly affect the rate constants for hydride transfers. In contrast, other studies show that proximal residues significantly participate in catalysis.

PubMed: 29271062
DOI: 10.1002/pro.3370

実験手法

X-RAY DIFFRACTION (1.2 Å)