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Literature summary extracted from

  • Dhagat, U.; Endo, S.; Mamiya, H.; Hara, A.; El-Kabbani, O.
    Studies on a Tyr residue critical for the binding of coenzyme and substrate in mouse 3(17)alpha-hydroxysteroid dehydrogenase (AKR1C21): structure of the Y224D mutant enzyme (2010), Acta Crystallogr. Sect. D, 66, 198-204.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.209 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

EC Number Protein Variants Comment Organism
1.1.1.209 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
1.1.1.209 Y224D mutation reduces the Km value for NADP(H) by up to 80fold and completely reverses the 17alpha stereospecificity of the enzyme Mus musculus
1.1.1.209 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.209 0.0001
-
 lithocholic acid wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0002
-
 4-androstene-3,17-dione wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0002
-
 5beta-pregnane-20alpha-ol-3-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0002
-
 lithocholic acid mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0004
-
 5alpha-Androstan-17alpha-ol-3-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0004
-
 5beta-pregnane-3,20-dione mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0005
-
 5beta-pregnane-3,20-dione mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0006
-
 5beta-pregnane-3,20-dione wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0006
-
 Epitestosterone wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0006
-
 5alpha-Androstan-3alpha-ol-17-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0006
-
 5beta-pregnane-3alpha,20alpha-diol wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0006
-
 lithocholic acid mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0006
-
 5beta-pregnane-20alpha-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0007
-
 5beta-pregnane-20alpha-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0015
-
 4-androstene-3,17-dione mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0018
-
 5alpha-Androstan-3alpha-ol-17-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0032
-
 5beta-pregnane-3alpha,20alpha-diol mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0034
-
 5alpha-Androstan-3alpha-ol-17-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0038
-
 5beta-pregnane-3alpha,20alpha-diol mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0047
-
 5alpha-androstan-17beta-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0054
-
 5alpha-Androstan-17alpha-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0059
-
 4-androstene-3,17-dione mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0059
-
 5beta-pregnane-3,20-dione mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0067
-
 lithocholic acid mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0078
-
 4-androstene-3,17-dione mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.013
-
 5alpha-androstan-3beta,17beta-diol mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.014
-
 5alpha-Androstan-17alpha-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.014
-
 Epitestosterone mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.015
-
 5alpha-Androstan-3alpha-ol-17-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.015
-
 5beta-pregnane-20alpha-ol-3-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.018
-
 5beta-pregnane-3alpha,20alpha-diol mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.02
-
 Epitestosterone mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.035
-
 5alpha-androstan-3beta,17beta-diol mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.08
-
 5alpha-androstan-17beta-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.096
-
 5alpha-androstan-17beta-ol-3-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.1
-
 5alpha-androstan-3beta,17beta-diol mutant Q222N, pH 7.0, 25°C Mus musculus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.209 Mus musculus Q91WR5
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.209 4-androstene-3,17-dione + NADPH + H+
-
 Mus musculus ? + NADP+
-
 r
1.1.1.209 5-androstene-3beta,17beta-diol + NADP+ no substrate for wild-type, but substrate for mutants Y224F, Y224D, Q222D Mus musculus ? + NADPH
-
 ?
1.1.1.209 5alpha-androstan-17alpha-ol-3-one + NADP+
-
 Mus musculus 5alpha-androstan-3,17-dione + NADPH + H+
-
 ?
1.1.1.209 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 ?
1.1.1.209 5alpha-androstan-3alpha-ol-17-one + NADPH + H+
-
 Mus musculus 5alpha-androstan-3alpha-ol-17beta-ol + NADP+
-
 ?
1.1.1.209 5alpha-androstan-3beta,17beta-diol + NADP+
-
 Mus musculus ? + NADPH
-
 ?
1.1.1.209 5alpha-androstan-3beta,17beta-diol + NADP+
-
 Mus musculus ? + NADPH + H+
-
 ?
1.1.1.209 5beta-pregnane-20alpha-ol-3-one + NADP+
-
 Mus musculus 5beta-pregnane-3alpha,20alpha-diol + NADPH + H+
-
 r
1.1.1.209 5beta-pregnane-3,20-dione + NADPH + H+
-
 Mus musculus 5beta-pregnane-3alpha-ol-20-one + NADP+
-
 r
1.1.1.209 5beta-pregnane-3alpha,20alpha-diol + NADPH + H+
-
 Mus musculus 5beta-pregnane-20alpha-ol-3-one + NADP+
-
 r
1.1.1.209 epitestosterone + NADP+
-
 Mus musculus testosterone + NADPH + H+
-
 ?
1.1.1.209 lithocholic acid + NADP+
-
 Mus musculus ? + NADPH + H+
-
 r

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.209 0.00167
-
 5alpha-androstan-3beta,17beta-diol mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.00217
-
 5alpha-androstan-3beta,17beta-diol mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0025
-
 5alpha-androstan-17beta-ol-3-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0027
-
 5alpha-androstan-17beta-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.00367
-
 5beta-pregnane-3alpha,20alpha-diol mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0073
-
 lithocholic acid mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0217
-
 4-androstene-3,17-dione mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0217
-
 5alpha-Androstan-3alpha-ol-17-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.0367
-
 5alpha-androstan-17beta-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.047
-
 lithocholic acid wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.065
-
 5alpha-androstan-3beta,17beta-diol mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.073
-
 5alpha-Androstan-3alpha-ol-17-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.081
-
 lithocholic acid mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.105
-
 lithocholic acid mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.127
-
 5alpha-Androstan-3alpha-ol-17-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.167
-
 4-androstene-3,17-dione wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.167
-
 5alpha-androstan-17beta-ol-3-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.167
-
 5alpha-Androstan-17alpha-ol-3-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.167
-
 5alpha-androstan-3beta,17beta-diol wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.217
-
 5beta-pregnane-3alpha,20alpha-diol wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.23
-
 Epitestosterone mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.233
-
 5alpha-Androstan-3alpha-ol-17-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.25
-
 5beta-pregnane-3alpha,20alpha-diol mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.283
-
 5alpha-Androstan-17alpha-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.3
-
 Epitestosterone wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.317
-
 4-androstene-3,17-dione mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.35
-
 5beta-pregnane-3,20-dione mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.383
-
 5beta-pregnane-20alpha-ol-3-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.4
-
 5beta-pregnane-3,20-dione wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 0.4
-
 5beta-pregnane-3alpha,20alpha-diol mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.57
-
 Epitestosterone mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.617
-
 4-androstene-3,17-dione mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 0.867
-
 5alpha-Androstan-17alpha-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 1.25
-
 5beta-pregnane-20alpha-ol-3-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 2.83
-
 5beta-pregnane-20alpha-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 4
-
 5beta-pregnane-3,20-dione mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 4.83
-
 5beta-pregnane-3,20-dione mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 4.83
-
 5beta-pregnane-20alpha-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.209 0.03
-
 5alpha-androstan-17beta-ol-3-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.05
-
 5alpha-androstan-3beta,17beta-diol mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 0.167
-
 5alpha-androstan-3beta,17beta-diol mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.5
-
 5alpha-androstan-17beta-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 0.65
-
 5alpha-androstan-3beta,17beta-diol mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 1
-
 lithocholic acid mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 1
-
 5beta-pregnane-3alpha,20alpha-diol mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 1.5
-
 5alpha-Androstan-3alpha-ol-17-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 3.3
-
 4-androstene-3,17-dione mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 4.5
-
 5alpha-androstan-17beta-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 11.67
-
 Epitestosterone mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 20
-
 5alpha-Androstan-17alpha-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 40
-
 Epitestosterone mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 60
-
 5beta-pregnane-3,20-dione mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 65
-
 5beta-pregnane-3alpha,20alpha-diol mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 68.3
-
 5alpha-Androstan-3alpha-ol-17-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 70
-
 5alpha-Androstan-3alpha-ol-17-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 83
-
 5beta-pregnane-20alpha-ol-3-one mutant Y224D, pH 7.0, 25°C Mus musculus
1.1.1.209 105
-
 4-androstene-3,17-dione mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 122
-
 5alpha-Androstan-3alpha-ol-17-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 125
-
 5beta-pregnane-3alpha,20alpha-diol mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 160
-
 5alpha-Androstan-17alpha-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 183
-
 lithocholic acid mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 217
-
 4-androstene-3,17-dione mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 366
-
 5beta-pregnane-3alpha,20alpha-diol wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 400
-
 5beta-pregnane-20alpha-ol-3-one mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 400
-
 lithocholic acid mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 417
-
 5alpha-Androstan-17alpha-ol-3-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 467
-
 lithocholic acid wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 500
-
 Epitestosterone wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 667
-
 5beta-pregnane-3,20-dione wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 833
-
 4-androstene-3,17-dione wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 1200
-
 5beta-pregnane-3,20-dione mutant Q222N, pH 7.0, 25°C Mus musculus
1.1.1.209 2000
-
 5beta-pregnane-20alpha-ol-3-one wild-type, pH 7.0, 25°C Mus musculus
1.1.1.209 8000
-
 5beta-pregnane-3,20-dione mutant Y224F, pH 7.0, 25°C Mus musculus
1.1.1.209 8000
-
 5beta-pregnane-20alpha-ol-3-one mutant Y224F, pH 7.0, 25°C Mus musculus