Cloned (Comment) | Organism |
---|---|
expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain JM107 | Curvularia lunata |
Protein Variants | Comment | Organism |
---|---|---|
H111A | site-directed mutagenesis, the mutant is dimeric like the wild-type enzyme, inactive mutant | Curvularia lunata |
H111L | site-directed mutagenesis, the mutation renders the enzyme monomeric, inactive mutant | Curvularia lunata |
R129D | site-directed mutagenesis, the mutation renders the enzyme monomeric, the mutant shows reduced activity compared to the wild-type enzyme | Curvularia lunata |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00006 | - |
NADP+ | pH 8.0, 25°C, wild-type enzyme | Curvularia lunata | |
0.0046 | - |
NADP+ | pH 8.0, 25°C, mutant H111A | Curvularia lunata | |
0.0065 | - |
NADPH | pH 8.0, 25°C, wild-type enzyme | Curvularia lunata | |
0.0506 | - |
NADPH | pH 8.0, 25°C, mutant H111A | Curvularia lunata |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-estrene-3,17-dione + NADPH | Curvularia lunata | - |
4-estrene-17beta-ol-3-one + NADP+ | - |
r | |
estrone + NADPH | Curvularia lunata | - |
17beta-estradiol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Curvularia lunata | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain JM107 by glutathione affinity chromatography | Curvularia lunata |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
17beta-estradiol + NAD(P)+ = estrone + NAD(P)H + H+ | structure-function analysis, dimerization is crucial for catalytic activity | Curvularia lunata |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-estrene-3,17-dione + NADPH | - |
Curvularia lunata | 4-estrene-17beta-ol-3-one + NADP+ | - |
r | |
estrone + NADPH | - |
Curvularia lunata | 17beta-estradiol + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | wild-type 17beta-HSD and mutant H111A | Curvularia lunata |
monomer | mutants R129D and H111L, the monomeric enzymes are catalytically inactive | Curvularia lunata |
More | the enzyme functions as dimer or tetramer, but is a dimer under native conditions, secondary structure analysis of wild-type and mutant enzymes, and structure-function analysis, residues involved in dimerization or oligomerization of the enzyme via salt bridges or hydrophobic interactions, overview | Curvularia lunata |
tetramer | - |
Curvularia lunata |
Synonyms | Comment | Organism |
---|---|---|
17beta-HSD | - |
Curvularia lunata |
More | the enzyme belongs to the short-chain dehydrogenase/reductase superfamily | Curvularia lunata |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Curvularia lunata |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
denaturation of monomeric enzyme at higher temperatures | Curvularia lunata |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.65 | - |
NADP+ | pH 8.0, 25°C, wild-type enzyme | Curvularia lunata | |
0.9 | - |
NADP+ | pH 8.0, 25°C, mutant H111A | Curvularia lunata | |
2.4 | - |
NADPH | pH 8.0, 25°C, mutant H111A | Curvularia lunata | |
3.8 | - |
NADPH | pH 8.0, 25°C, wild-type enzyme | Curvularia lunata |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Curvularia lunata |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | dependent on | Curvularia lunata | |
NADPH | dependent on | Curvularia lunata |