Crystallization (Comment) | Organism |
---|---|
10 mg/ml purified double mutant H51Q/K228R in complex with NAD+ and 2,3- or 2,4-difluorobenzyl alcohol, in 50 mM ammonium N-[tris(hydroxymethyl)-methyl]-2-aminoethanesulfonate, pH 7.0, 5°C, 1 mM NAD+, 10 mM 2,3-difluorobenzyl alcohol or 2,4-difluorobenzyl alcohol, equilibrated against increasing concentrations of 2-methyl-2,4-pentanediole, crystal formation at 12% 2-methyl-2,4-pentanediole, X-ray diffraction structure determination and analysis | Equus caballus |
Protein Variants | Comment | Organism |
---|---|---|
H51Q | site-directed mutagenesis, shifting of pH dependency, increased activity at pH 8.0, decrease of the rate of isomerization of the enzyme-NAD+ complex, which becomes the limiting step for ethanol oxidation | Equus caballus |
H51Q/K228R | site-directed mutagenesis, kinetic effects | Equus caballus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | detailed kinetic mechanism, steady-state kinetics for wild-type and mutant enzymes, investigation of pH-dependency | Equus caballus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | catalytic zinc ion | Equus caballus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Equus caballus | P00327 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
a primary alcohol + NAD+ = an aldehyde + NADH + H+ | reaction mechanism, His51 is involved, but not essential, in catalysis facilitating the deprotonation of the hydroxyl group of water or alcohol ligated to the catalytic zinc | Equus caballus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Equus caballus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
butanol + NAD+ | - |
Equus caballus | n-butanal + NADH | - |
? | |
ethanol + NAD+ | isomerization of the enzyme-NAD+ complex is the rate-limiting step for acetaldehyde reduction by the wild-type enzyme | Equus caballus | acetaldehyde + NADH | - |
r | |
n-propanol + NAD+ | - |
Equus caballus | propanal + NADH | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ADH | - |
Equus caballus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Equus caballus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 10 | - |
Equus caballus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Equus caballus |