5KJE: F322L horse liver alcohol dehydrogenase complexed with NAD+ and pentafluorobenzyl alcohol

Citation:
Abstract
The dynamics of enzyme catalysis range from the slow time scale ( approximately ms) for substrate binding and conformational changes to the fast time ( approximately 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.
PDB ID: 5KJEDownload
MMDB ID: 141073
PDB Deposition Date: 2016/6/18
Updated in MMDB: 2017/10
Experimental Method:
x-ray diffraction
Resolution: 1.26  Å
Source Organism:
Similar Structures:
Biological Unit for 5KJE: dimeric; determined by author and by software (PISA)
Molecular Components in 5KJE
Label Count Molecule
Proteins (2 molecules)
2
Alcohol Dehydrogenase E Chain(Gene symbol: LOC100034242)
Molecule annotation
Chemicals (12 molecules)
1
4
2
2
3
2
4
4
* Click molecule labels to explore molecular sequence information.

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