Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
H146L | site-directed mutagenesis, altered activity | Rattus norvegicus |
K153M | site-directed mutagenesis, active site mutant, complete loss of activity | Rattus norvegicus |
L143F | site-directed mutagenesis, altered activity | Rattus norvegicus |
N190V | site-directed mutagenesis, altered activity | Rattus norvegicus |
N190V/W191S | site-directed mutagenesis, almost complete loss of L-xylulose reductase activity | Rattus norvegicus |
N190V/W191S/Q137M/L143F/H146L | site-directed mutagenesis, almost complete loss of L-xylulose reductase activity, mutant shows high 3-ketosteroid reductase activity | Rattus norvegicus |
Q137M | site-directed mutagenesis, altered activity, stable against cold inactivation | Rattus norvegicus |
Q137M/F241L | site-directed mutagenesis, altered activity, sensitive to cold inactivation like the wild-type enzyme | Rattus norvegicus |
Q137M/L143F | site-directed mutagenesis, increased Km for L-xylulose compared to the wild-type | Rattus norvegicus |
Q137M/L143F/H146L | site-directed mutagenesis, almost complete loss of L-xylulose reductase activity, mutant shows 3-ketosteroid reductase activity | Rattus norvegicus |
S136A | site-directed mutagenesis, active site mutant, complete loss of activity | Rattus norvegicus |
W191F | site-directed mutagenesis, altered activity | Rattus norvegicus |
W191S | site-directed mutagenesis, altered activity | Rattus norvegicus |
Y149F | site-directed mutagenesis, active site mutant, complete loss of activity | Rattus norvegicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
n-butyric acid | specific, binds to the enzyme-NADP+ complex, the mutant enzymes show altered sensitivity to inhibition, overview | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00067 | - |
NADP+ | pH 7.0, 25°C, mutant S136A | Rattus norvegicus | |
0.0013 | - |
NADP+ | pH 7.0, 25°C, mutant Y149F | Rattus norvegicus | |
0.002 | - |
NADPH | pH 7.0, 25°C, mutant Y149F | Rattus norvegicus | |
0.003 | - |
NADP+ | pH 7.0, 25°C, wild-type enzyme | Rattus norvegicus | |
0.0046 | - |
NADPH | pH 7.0, 25°C, mutant S136A | Rattus norvegicus | |
0.005 | - |
1,4-dibromo-2,3-butanedione | pH 7.0, 25°C, wild-type enzyme | Rattus norvegicus | |
0.007 | - |
NADPH | pH 7.0, 25°C, wild-type enzyme | Rattus norvegicus | |
0.0099 | - |
L-xylitol | pH 7.0, 25°C, mutant Y149F | Rattus norvegicus | |
0.011 | - |
L-xylitol | pH 7.0, 25°C, wild-type enzyme | Rattus norvegicus | |
0.013 | - |
L-xylitol | pH 7.0, 25°C, mutant S136A | Rattus norvegicus | |
0.05 | - |
1,4-dibromo-2,3-butanedione | pH 7.0, 25°C, mutant S136A | Rattus norvegicus | |
0.053 | - |
NADPH | pH 7.0, 25°C, mutant K153M | Rattus norvegicus | |
0.085 | - |
L-xylitol | pH 7.0, 25°C, mutant K153M | Rattus norvegicus | |
0.092 | - |
L-xylulose | pH 7.0, 25°C, mutant H146L | Rattus norvegicus | |
0.1 | - |
1,4-dibromo-2,3-butanedione | pH 7.0, 25°C, mutant K153M | Rattus norvegicus | |
0.117 | - |
NADP+ | pH 7.0, 25°C, mutant K153M | Rattus norvegicus | |
0.139 | - |
1,4-dibromo-2,3-butanedione | pH 7.0, 25°C, mutant Y149F | Rattus norvegicus | |
0.14 | - |
L-xylulose | pH 7.0, 25°C, wild-type enzyme and mutant L143F | Rattus norvegicus | |
0.31 | - |
L-xylulose | pH 7.0, 25°C, mutant N190V | Rattus norvegicus | |
0.33 | - |
L-xylulose | pH 7.0, 25°C, mutant Q137M | Rattus norvegicus | |
0.78 | - |
diacetyl | pH 7.0, 25°C, wild-type enzyme | Rattus norvegicus | |
1.2 | - |
diacetyl | pH 7.0, 25°C, mutant H146L | Rattus norvegicus | |
2.6 | - |
diacetyl | pH 7.0, 25°C, mutant N190V | Rattus norvegicus | |
2.7 | - |
diacetyl | pH 7.0, 25°C, mutant N190V/W191S/Q137M/L143F/H146L | Rattus norvegicus | |
3 | - |
diacetyl | pH 7.0, 25°C, mutant Q137M | Rattus norvegicus | |
3.6 | - |
L-xylulose | pH 7.0, 25°C, mutant Q137M/L143F | Rattus norvegicus | |
5.2 | - |
diacetyl | pH 7.0, 25°C, mutant W191F | Rattus norvegicus | |
6.2 | - |
diacetyl | pH 7.0, 25°C, mutant L143F | Rattus norvegicus | |
7.2 | - |
L-xylulose | pH 7.0, 25°C, mutant W191F | Rattus norvegicus | |
9.6 | - |
diacetyl | pH 7.0, 25°C, mutant Q137M/L143F/H146L | Rattus norvegicus | |
11 | - |
diacetyl | pH 7.0, 25°C, mutant Q137M/L143F | Rattus norvegicus | |
16 | - |
diacetyl | pH 7.0, 25°C, mutant N190V/W191S | Rattus norvegicus | |
42 | - |
diacetyl | pH 7.0, 25°C, mutant W191S | Rattus norvegicus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | - |
Rattus norvegicus | 16020 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
26500 | - |
x * 26500 | Rattus norvegicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
diacetyl + NAD(P)H | Rattus norvegicus | detoxification of alpha-dicarbonyl compounds | acetoin + NAD(P)+ | - |
r | |
L-xylulose + NADPH + H+ | Rattus norvegicus | part of the uronate cycle, involved in osmoregulation in the kidney | L-xylitol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
xylitol + NADP+ = L-xylulose + NADPH + H+ | enzyme with dual function showing L-xylulose reductase and dicarbonyl reductase activities, it is probably identical with sperm 34 kDa protein P34H and diacetyl reductase, EC 1.1.1.5, the amino acid residues Ser136, Tyr149, and Lys153 form the catalytic triad | Rattus norvegicus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
kidney | brush border | Rattus norvegicus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,4-dibromo-2,3-butanedione + NADPH | dicarbonyl reductase activity | Rattus norvegicus | ? + NADP+ | - |
r | |
diacetyl + NAD(P)H | detoxification of alpha-dicarbonyl compounds | Rattus norvegicus | acetoin + NAD(P)+ | - |
r | |
diacetyl + NAD(P)H | dicarbonyl reductase activity | Rattus norvegicus | acetoin + NAD(P)+ | - |
r | |
L-xylulose + NADPH + H+ | part of the uronate cycle, involved in osmoregulation in the kidney | Rattus norvegicus | L-xylitol + NADP+ | - |
r | |
L-xylulose + NADPH + H+ | L-xylulose reductase activity | Rattus norvegicus | L-xylitol + NADP+ | - |
r | |
additional information | the size and hydrophobicity of the amino acid residues involved in substrate recognition, i.e. Q137, L143, H146, N190, and W191, is important, mutants N190V, N190V/W191S, Q137M/L143F/H146L, and N190V/W191S/Q137M/L143F/H146L show reductive activity with 4-nitroacetophenone, 5beta-androstane-3,17-dione, 5beta-androstan-17beta-ol-3-one, and 5beta-androstane-3alpha,17beta-diol, overview | Rattus norvegicus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 26500 | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
More | enzyme belongs to the short-chain dehydrogenase/reductase family | Rattus norvegicus |
XR | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0005 | - |
NADPH | pH 7.0, 25°C, mutant K153M | Rattus norvegicus | |
0.0006 | - |
NADPH | pH 7.0, 25°C, mutant S136A | Rattus norvegicus | |
0.0009 | - |
NADPH | pH 7.0, 25°C, mutant Y149F | Rattus norvegicus | |
9.5 | - |
NADP+ | pH 7.0, 25°C, wild-type enzyme | Rattus norvegicus | |
25 | - |
NADPH | pH 7.0, 25°C, wild-type enzyme | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Rattus norvegicus | |
NADPH | - |
Rattus norvegicus |