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Literature summary for 1.1.1.209 extracted from

  • Dhagat, U.; Endo, S.; Mamiya, H.; Hara, A.; El-Kabbani, O.
    Structure of the G225P/G226P mutant of mouse 3(17)alpha-hydroxysteroid dehydrogenase (AKR1C21) ternary complex: implications for the binding of inhibitor and substrate (2009), Acta Crystallogr. Sect. D, 65, 257-265.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant wild-type and mutant enzymes expressed in Escherichia coli JM109 Mus musculus

Crystallization (Commentary)

Crystallization (Comment) Organism
double mutant G225P/G226P of AKR1C21 in complex with the coenzyme NADP+ and the inhibitor hexoestrol, by hanging-drop vapour-diffusion method, at 2.1 A resolution. Overall fold of the mutant is similar to that of wild-type, apart from the loop region lining the active-site pocket, in which the two glycines at positions 225 and 226 are mutated to prolines which have a more rigid structure, resulting in a larger substrate-binding pocket. The mutation affects the conformation of the adjoining residues Tyr224 and Trp227, resulting in a further opening up of the active-site pocket while the catalytic residues remain unaffected. The inhibitor molecule is bound to the active site with one hydroxyl group pointing towards the catalytic residues, the bulky phenyl rings and the methyl groups of hexoestrol form van der Waals contacts with Trp227, Tyr224, Tyr55 and the nicotinamide ring of NADP+ Mus musculus

Protein Variants

Protein Variants Comment Organism
G225P/G226P reduced affinity (Km) for both 3alpha- and 17alpha-hydroxysteroid substrates by up to 160fold, shows high selectivity for 5beta-pregnane-3alpha,20alpha-diol in the oxidation of the 3alpha-hydroxy group and, conversely, broad specificity for both 17alpha- and 17beta-hydroxysteroids Mus musculus

Inhibitors

Inhibitors Comment Organism Structure
Hexoestrol
-
Mus musculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0002
-
4-androstene-3,17-dione wild-type Mus musculus
0.0004
-
5alpha-Androstan-17alpha-ol-3-one wild-type Mus musculus
0.0004
-
5beta-androstan-17alpha-ol-3-one wild-type Mus musculus
0.0005
-
5alpha-pregnane-3alpha,20alpha-diol wild-type Mus musculus
0.0006
-
Epitestosterone wild-type Mus musculus
0.0006
-
5alpha-Androstan-3alpha-ol-17-one wild-type Mus musculus
0.0006
-
5beta-pregnane-3alpha,20alpha-diol wild-type Mus musculus
0.0009
-
4-androstene-3,17-dione mutant G225P/G226P Mus musculus
0.0013
-
5beta-androstan-17alpha-ol-3-one mutant G225P/G226P Mus musculus
0.0029
-
5beta-pregnane-3alpha,20alpha-diol mutant G225P/G226P Mus musculus
0.0048
-
5alpha-Androstan-17alpha-ol-3-one mutant G225P/G226P Mus musculus
0.0097
-
5alpha-Androstan-3alpha-ol-17-one mutant G225P/G226P Mus musculus
0.018
-
Epitestosterone mutant G225P/G226P Mus musculus
0.023
-
testosterone mutant G225P/G226P Mus musculus
0.027
-
5alpha-androstan-17beta-ol-3-one mutant G225P/G226P Mus musculus
0.08
-
5alpha-pregnane-3alpha,20alpha-diol mutant G225P/G226P Mus musculus

Organism

Organism UniProt Comment Textmining
Mus musculus Q91WR5
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes purified to homogeneity Mus musculus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-androstene-3,17-dione + NADPH
-
Mus musculus ? + NADP+
-
?
5alpha-androstan-17alpha-ol-3-one + NADP+
-
Mus musculus ? + NADPH
-
?
5alpha-androstan-17beta-ol-3-one + NADP+ wild-type shows no activity with 5alpha-androstan-17beta-ol-3-one Mus musculus 5alpha-androstan-3,17-dione + NADPH + H+
-
?
5alpha-androstan-3alpha-ol-17-one + NADPH
-
Mus musculus ? + NADP+
-
?
5alpha-pregnane-3alpha,20alpha-diol + NADP+
-
Mus musculus ? + NADPH
-
?
5beta-androstan-17alpha-ol-3-one + NADP+
-
Mus musculus ? + NADPH
-
?
5beta-pregnane-3alpha,20alpha-diol + NADP+
-
Mus musculus ? + NADPH
-
?
epitestosterone
-
Mus musculus ? + NADPH
-
?
additional information wild-type shows no activity with testosterone and 5alpha-androstan-17beta-ol-3-one Mus musculus ?
-
?
testosterone + NADP+ mutant G225P/G226P, wild-type shows no activity with testosterone Mus musculus androst-4-en-3,17-dione + NADPH + H+
-
?

Synonyms

Synonyms Comment Organism
3(17)alpha-hydroxysteroid dehydrogenase
-
Mus musculus
AKR1C21
-
Mus musculus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.04
-
testosterone mutant G225P/G226P Mus musculus
0.055
-
5beta-androstan-17alpha-ol-3-one mutant G225P/G226P Mus musculus
0.073
-
5alpha-Androstan-3alpha-ol-17-one wild-type Mus musculus
0.105
-
5alpha-pregnane-3alpha,20alpha-diol wild-type Mus musculus
0.1367
-
4-androstene-3,17-dione mutant G225P/G226P Mus musculus
0.14
-
5beta-androstan-17alpha-ol-3-one wild-type Mus musculus
0.145
-
5alpha-Androstan-17alpha-ol-3-one mutant G225P/G226P Mus musculus
0.15
-
5alpha-pregnane-3alpha,20alpha-diol mutant G225P/G226P Mus musculus
0.167
-
4-androstene-3,17-dione wild-type Mus musculus
0.167
-
5alpha-Androstan-17alpha-ol-3-one wild-type Mus musculus
0.183
-
5beta-pregnane-3alpha,20alpha-diol mutant G225P/G226P Mus musculus
0.2167
-
5beta-pregnane-3alpha,20alpha-diol wild-type Mus musculus
0.2167
-
5alpha-Androstan-3alpha-ol-17-one mutant G225P/G226P Mus musculus
0.25
-
Epitestosterone mutant G225P/G226P Mus musculus
0.3
-
Epitestosterone wild-type Mus musculus
0.383
-
5alpha-androstan-17beta-ol-3-one mutant G225P/G226P Mus musculus

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Mus musculus

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00026
-
wild-type Mus musculus Hexoestrol
0.0006
-
mutant G225P/G226P Mus musculus Hexoestrol