Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant isozyme type-2 in Escherichia coli strain Bl21(DE3) | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
recombinant isozyme type-2 mutant L459F/N460G in complex with NAD+ and in absence of substrate, hanging drop vapour diffusion method, 7.6 mg/ml enzyme with 2 mM NAD+ and about 14 mg/ml retinol, 0.002 ml protein solution plus equal volume of well solution containing 7% PEG 6000, 2% 2-methyl-2,4-pentanediol, and 0.1 M Tris, pH 7.5, X-ray diffraction structure determination and analysis at 3.3 A resolution | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
C301A | site-directed mutagenesis, catalytic residue mutation, reduced activity compared to the wild-type enzyme | Mus musculus |
L459F/N460G | site-directed mutagenesis, inactive mutant, still able to bind the cofactor NAD+ | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
decanal | recombinant wild-type enzyme and mutant L459F/N460G, pH 8.5, 25°C | Mus musculus | |
0.005 | - |
octanal | recombinant mutant L459F/N460G, pH 8.5, 25°C | Mus musculus | |
0.005 | - |
hexanal | recombinant wild-type enzyme, pH 8.5, 25°C | Mus musculus | |
0.008 | - |
hexanal | recombinant mutant L459F/N460G, pH 8.5, 25°C | Mus musculus | |
0.01 | - |
octanal | recombinant wild-type enzyme, pH 8.5, 25°C | Mus musculus | |
0.014 | - |
acetaldehyde | recombinant wild-type enzyme, pH 8.5, 25°C | Mus musculus | |
0.031 | - |
acetaldehyde | recombinant mutant L459F/N460G, pH 8.5, 25°C | Mus musculus | |
0.067 | - |
NAD+ | recombinant wild-type enzyme, pH 8.5, 25°C | Mus musculus | |
0.39 | - |
NAD+ | recombinant mutant L459F/N460G, pH 8.5, 25°C | Mus musculus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | - |
isozyme type-2 | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant isozyme type-2 from Escherichia coli strain Bl21(DE3), no usage of His-tag since it may influence the enzyme activity | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetaldehyde + NAD+ + H2O | - |
Mus musculus | acetate + NADH + H+ | - |
? | |
decanal + NAD+ + H2O | - |
Mus musculus | decanoate + NADH | - |
? | |
hexanal + NAD+ + H2O | - |
Mus musculus | hexanoate + NADH + H+ | - |
? | |
additional information | substrate specificities of wild-type isozyme type-2 and mutant L459F/N460G | Mus musculus | ? | - |
? | |
octanal + NAD+ + H2O | - |
Mus musculus | octanoate + NADH | - |
? |
Synonyms | Comment | Organism |
---|---|---|
RALDH2 | - |
Mus musculus |
retinaldehyde dehydrogenase type II | - |
Mus musculus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Mus musculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Mus musculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Mus musculus |