Crystallization (Comment) | Organism |
---|---|
Y224D mutant at 2.3 A resolution. Mutation results in a change in the conformation of the flexible loop B, including the V-shaped groove | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
Q222N | mutation of residue involved in forming the safety belt for binding of the coenzyme. Kinetic constants for 3-hydroxy/3-ketosteroids and 17-hydroxy/ketosteroids similar to wild type | Mus musculus |
Y224D | mutation reduces the Km value for NADP(H) by up to 80fold and completely reverses the 17alpha stereospecificity of the enzyme | Mus musculus |
Y224F | mutation of residue involved in forming the safety belt for binding of the coenzyme. Kinetic constants for 3-hydroxy/3-ketosteroids and 17-hydroxy/ketosteroids similar to wild type | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0001 | - |
lithocholic acid | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0002 | - |
4-androstene-3,17-dione | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0002 | - |
5beta-pregnane-20alpha-ol-3-one | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0002 | - |
lithocholic acid | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0004 | - |
5alpha-Androstan-17alpha-ol-3-one | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0004 | - |
5beta-pregnane-3,20-dione | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.0005 | - |
5beta-pregnane-3,20-dione | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0006 | - |
5beta-pregnane-3,20-dione | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0006 | - |
Epitestosterone | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0006 | - |
5alpha-Androstan-3alpha-ol-17-one | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0006 | - |
5beta-pregnane-3alpha,20alpha-diol | wild-type, pH 7.0, 25°C | Mus musculus | |
0.0006 | - |
lithocholic acid | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.0006 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0007 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.0015 | - |
4-androstene-3,17-dione | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0018 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0032 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0034 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.0038 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.0047 | - |
5alpha-androstan-17beta-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0054 | - |
5alpha-Androstan-17alpha-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0059 | - |
4-androstene-3,17-dione | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.0059 | - |
5beta-pregnane-3,20-dione | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.0067 | - |
lithocholic acid | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.0078 | - |
4-androstene-3,17-dione | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.013 | - |
5alpha-androstan-3beta,17beta-diol | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.014 | - |
5alpha-Androstan-17alpha-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.014 | - |
Epitestosterone | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.015 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.015 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.018 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.02 | - |
Epitestosterone | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.035 | - |
5alpha-androstan-3beta,17beta-diol | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.08 | - |
5alpha-androstan-17beta-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.096 | - |
5alpha-androstan-17beta-ol-3-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.1 | - |
5alpha-androstan-3beta,17beta-diol | mutant Q222N, pH 7.0, 25°C | Mus musculus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | Q91WR5 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-androstene-3,17-dione + NADPH + H+ | - |
Mus musculus | ? + NADP+ | - |
r | |
5-androstene-3beta,17beta-diol + NADP+ | no substrate for wild-type, but substrate for mutants Y224F, Y224D, Q222D | Mus musculus | ? + NADPH | - |
? | |
5alpha-androstan-17alpha-ol-3-one + NADP+ | - |
Mus musculus | 5alpha-androstan-3,17-dione + NADPH + H+ | - |
? | |
5alpha-androstan-17beta-ol-3-one + NADP+ | - |
Mus musculus | 5alpha-androstan-3,17-dione + NADPH + H+ | no substrate for wild-type, but substrate for mutants Y224F, Y224D, Q222D | ? | |
5alpha-androstan-3alpha-ol-17-one + NADPH + H+ | - |
Mus musculus | 5alpha-androstan-3alpha-ol-17beta-ol + NADP+ | - |
? | |
5alpha-androstan-3beta,17beta-diol + NADP+ | - |
Mus musculus | ? + NADPH | - |
? | |
5alpha-androstan-3beta,17beta-diol + NADP+ | - |
Mus musculus | ? + NADPH + H+ | - |
? | |
5beta-pregnane-20alpha-ol-3-one + NADP+ | - |
Mus musculus | 5beta-pregnane-3alpha,20alpha-diol + NADPH + H+ | - |
r | |
5beta-pregnane-3,20-dione + NADPH + H+ | - |
Mus musculus | 5beta-pregnane-3alpha-ol-20-one + NADP+ | - |
r | |
5beta-pregnane-3alpha,20alpha-diol + NADPH + H+ | - |
Mus musculus | 5beta-pregnane-20alpha-ol-3-one + NADP+ | - |
r | |
epitestosterone + NADP+ | - |
Mus musculus | testosterone + NADPH + H+ | - |
? | |
lithocholic acid + NADP+ | - |
Mus musculus | ? + NADPH + H+ | - |
r |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00167 | - |
5alpha-androstan-3beta,17beta-diol | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.00217 | - |
5alpha-androstan-3beta,17beta-diol | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.0025 | - |
5alpha-androstan-17beta-ol-3-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.0027 | - |
5alpha-androstan-17beta-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.00367 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.0073 | - |
lithocholic acid | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.0217 | - |
4-androstene-3,17-dione | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.0217 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.0367 | - |
5alpha-androstan-17beta-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.047 | - |
lithocholic acid | wild-type, pH 7.0, 25°C | Mus musculus | |
0.065 | - |
5alpha-androstan-3beta,17beta-diol | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.073 | - |
5alpha-Androstan-3alpha-ol-17-one | wild-type, pH 7.0, 25°C | Mus musculus | |
0.081 | - |
lithocholic acid | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.105 | - |
lithocholic acid | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.127 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.167 | - |
4-androstene-3,17-dione | wild-type, pH 7.0, 25°C | Mus musculus | |
0.167 | - |
5alpha-androstan-17beta-ol-3-one | wild-type, pH 7.0, 25°C | Mus musculus | |
0.167 | - |
5alpha-Androstan-17alpha-ol-3-one | wild-type, pH 7.0, 25°C | Mus musculus | |
0.167 | - |
5alpha-androstan-3beta,17beta-diol | wild-type, pH 7.0, 25°C | Mus musculus | |
0.217 | - |
5beta-pregnane-3alpha,20alpha-diol | wild-type, pH 7.0, 25°C | Mus musculus | |
0.23 | - |
Epitestosterone | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.233 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.25 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.283 | - |
5alpha-Androstan-17alpha-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.3 | - |
Epitestosterone | wild-type, pH 7.0, 25°C | Mus musculus | |
0.317 | - |
4-androstene-3,17-dione | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.35 | - |
5beta-pregnane-3,20-dione | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.383 | - |
5beta-pregnane-20alpha-ol-3-one | wild-type, pH 7.0, 25°C | Mus musculus | |
0.4 | - |
5beta-pregnane-3,20-dione | wild-type, pH 7.0, 25°C | Mus musculus | |
0.4 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.57 | - |
Epitestosterone | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.617 | - |
4-androstene-3,17-dione | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
0.867 | - |
5alpha-Androstan-17alpha-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
1.25 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
2.83 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
4 | - |
5beta-pregnane-3,20-dione | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
4.83 | - |
5beta-pregnane-3,20-dione | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
4.83 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
5alpha-androstan-17beta-ol-3-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.05 | - |
5alpha-androstan-3beta,17beta-diol | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
0.167 | - |
5alpha-androstan-3beta,17beta-diol | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.5 | - |
5alpha-androstan-17beta-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
0.65 | - |
5alpha-androstan-3beta,17beta-diol | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
1 | - |
lithocholic acid | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
1 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
1.5 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
3.3 | - |
4-androstene-3,17-dione | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
4.5 | - |
5alpha-androstan-17beta-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
11.67 | - |
Epitestosterone | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
20 | - |
5alpha-Androstan-17alpha-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
40 | - |
Epitestosterone | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
60 | - |
5beta-pregnane-3,20-dione | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
65 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
68.3 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
70 | - |
5alpha-Androstan-3alpha-ol-17-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
83 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Y224D, pH 7.0, 25°C | Mus musculus | |
105 | - |
4-androstene-3,17-dione | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
122 | - |
5alpha-Androstan-3alpha-ol-17-one | wild-type, pH 7.0, 25°C | Mus musculus | |
125 | - |
5beta-pregnane-3alpha,20alpha-diol | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
160 | - |
5alpha-Androstan-17alpha-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
183 | - |
lithocholic acid | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
217 | - |
4-androstene-3,17-dione | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
366 | - |
5beta-pregnane-3alpha,20alpha-diol | wild-type, pH 7.0, 25°C | Mus musculus | |
400 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
400 | - |
lithocholic acid | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
417 | - |
5alpha-Androstan-17alpha-ol-3-one | wild-type, pH 7.0, 25°C | Mus musculus | |
467 | - |
lithocholic acid | wild-type, pH 7.0, 25°C | Mus musculus | |
500 | - |
Epitestosterone | wild-type, pH 7.0, 25°C | Mus musculus | |
667 | - |
5beta-pregnane-3,20-dione | wild-type, pH 7.0, 25°C | Mus musculus | |
833 | - |
4-androstene-3,17-dione | wild-type, pH 7.0, 25°C | Mus musculus | |
1200 | - |
5beta-pregnane-3,20-dione | mutant Q222N, pH 7.0, 25°C | Mus musculus | |
2000 | - |
5beta-pregnane-20alpha-ol-3-one | wild-type, pH 7.0, 25°C | Mus musculus | |
8000 | - |
5beta-pregnane-3,20-dione | mutant Y224F, pH 7.0, 25°C | Mus musculus | |
8000 | - |
5beta-pregnane-20alpha-ol-3-one | mutant Y224F, pH 7.0, 25°C | Mus musculus |