5KJF

V222I horse liver alcohol dehydrogenase complexed with NAD+ and trifluoroethanol

Summary for 5KJF

• Entry DOI: 10.2210/pdb5kjf/pdb
• Related: 4DXH 5KCP 5KCZ 5KJ1 5KJ6 5KJC 5KJD
• Descriptor: Alcohol dehydrogenase E chain, ZINC ION, NICOTINAMIDE-ADENINE-DINUCLEOTIDE (ACIDIC FORM), ... (6 entities in total)
• Functional Keywords: oxidoreductase, rossmann fold, michaelis analogue, dynamics
• Biological source: Equus caballus (Horse)
• Total number of polymer chains: 2
• Total formula weight: 82114.04

Authors

Plapp, B.V. (deposition date: 2016-06-18, release date: 2016-07-27, Last modification date: 2023-09-27)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.2 Å)