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Literature summary for 1.1.1.201 extracted from

  • Savino, S.; Ferrandi, E.E.; Forneris, F.; Rovida, S.; Riva, S.; Monti, D.; Mattevi, A.
    Structural and biochemical insights into 7beta-hydroxysteroid dehydrogenase stereoselectivity (2016), Proteins, 84, 859-865.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Collinsella aerofaciens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified His-tagged enzyme complexed with NADP+, mixing of 10-15 mg/ml protein in 20% v/v glycerol, 20 mM potassium phosphate, pH 7.0, and 5 mM NADP+, with1 M sodium formate, 100 mM sodium acetate, pH 4.6, at room temperature, 3-4days, X-ray diffraction structure determination and analysis at 1.6 A resolution, modelling, molecular replacement using the the apoform structure of 7alpha-HSDH from Escherichia coli as search model, PDB ID 1AHH Collinsella aerofaciens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7beta-hydroxysteroid + NADP+ Collinsella aerofaciens
-
7-oxosteroid + NADPH + H+
-
r
7beta-hydroxysteroid + NADP+ Collinsella aerofaciens ATCC 25986
-
7-oxosteroid + NADPH + H+
-
r
ursodeoxycholic acid + NADP+ Collinsella aerofaciens
-
7-oxo-lithocholic acid + NADPH + H+
-
r
ursodeoxycholic acid + NADP+ Collinsella aerofaciens ATCC 25986
-
7-oxo-lithocholic acid + NADPH + H+
-
r

Organism

Organism UniProt Comment Textmining
Collinsella aerofaciens A4ECA9
-
-
Collinsella aerofaciens ATCC 25986 A4ECA9
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration Collinsella aerofaciens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ i.e. ursodeoxycholic acid, exclusive specificity for the hydroxyl group in beta configuration at the C-7 position Collinsella aerofaciens 7-oxo-lithocholic acid + NADPH + H+
-
r
3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ i.e. ursodeoxycholic acid, exclusive specificity for the hydroxyl group in beta configuration at the C-7 position Collinsella aerofaciens ATCC 25986 7-oxo-lithocholic acid + NADPH + H+
-
r
7beta-hydroxysteroid + NADP+
-
Collinsella aerofaciens 7-oxosteroid + NADPH + H+
-
r
7beta-hydroxysteroid + NADP+ stereoselective with 7-beta-hydroxysteroids Collinsella aerofaciens 7-oxosteroid + NADPH + H+
-
r
7beta-hydroxysteroid + NADP+
-
Collinsella aerofaciens ATCC 25986 7-oxosteroid + NADPH + H+
-
r
7beta-hydroxysteroid + NADP+ stereoselective with 7-beta-hydroxysteroids Collinsella aerofaciens ATCC 25986 7-oxosteroid + NADPH + H+
-
r
additional information no activity with chenodeoxycholic acid, i.e. 3alpha,7alpha-dihydroxy-5beta-cholan-24-oic acid Collinsella aerofaciens ?
-
?
additional information no activity with chenodeoxycholic acid, i.e. 3alpha,7alpha-dihydroxy-5beta-cholan-24-oic acid Collinsella aerofaciens ATCC 25986 ?
-
?
ursodeoxycholic acid + NADP+
-
Collinsella aerofaciens 7-oxo-lithocholic acid + NADPH + H+
-
r
ursodeoxycholic acid + NADP+
-
Collinsella aerofaciens ATCC 25986 7-oxo-lithocholic acid + NADPH + H+
-
r

Subunits

Subunits Comment Organism
homodimer a dimer of identical subunits arranged with C2 symmetry Collinsella aerofaciens
More crystal structure analysis reveals a loop gating active site accessibility, the bases of the specificity for NADP+, and the general architecture of the steroid binding site Collinsella aerofaciens

Synonyms

Synonyms Comment Organism
7beta-HSDH
-
Collinsella aerofaciens
7beta-hydroxysteroid dehydrogenase
-
Collinsella aerofaciens

Cofactor

Cofactor Comment Organism Structure
NADP+ binding site structure analysis Collinsella aerofaciens
NADPH
-
Collinsella aerofaciens

General Information

General Information Comment Organism
additional information crystal structure analysis reveals a loop gating active site accessibility, the bases of the specificity for NADP+, and the general architecture of the steroid binding site. Conserved residues Ser143 and Tyr156 of 7beta-HSDH are homologous with and structurally superimpose to Ser146 and Tyr159 of 7alpha-HSDH Collinsella aerofaciens