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1.14.14.23: cholesterol 7alpha-monooxygenase

This is an abbreviated version!
For detailed information about cholesterol 7alpha-monooxygenase, go to the full flat file.

Word Map on EC 1.14.14.23

Reaction

cholesterol
+
[reduced NADPH-hemoprotein reductase]
 +
O2
=
7alpha-hydroxycholesterol
 +
[oxidized NADPH-hemoprotein reductase]
 +
H2O

Synonyms

7alpha-hydroxylase, bile acid-synthetic enzyme, C7alphaOH, cholesterol 7 alpha-hydroxylase, cholesterol 7-alpha hydroxylase, Cholesterol 7-alpha-hydroxylase, Cholesterol 7-alpha-monooxygenase, cholesterol 7a-hydroxylase, cholesterol 7alpha hydroxylase, cholesterol 7alpha-hydroxylase, cholesterol-NADPH oxidoreductase, 7alpha-hydroxylating, CYP125A4, CYP7A, CYP7A1, Cyp7alpha1, CYPVII, cytochrome P450 125A4, EC 1.14.13.17, hepatic cholesterol 7alpha-hydroxylase, More, oxygenase, cholesterol 7alpha-mono-

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.14 With reduced flavin or flavoprotein as one donor, and incorporation of one atom of oxygen into the other donor
                1.14.14.23 cholesterol 7alpha-monooxygenase

Crystallization

Crystallization on EC 1.14.14.23 - cholesterol 7alpha-monooxygenase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
wild-type CYP7A1 in the ligand-free state, structure-based mutation T104L in the B' helix, corresponding to the nonpolar residue of CYP7B1, is used to obtain crystals of complexes with cholest-4-en-3 -one and with cholesterol oxidation product 7-ketocholesterol, hanging drop vapor diffusion method, mixing 0f 0.001 ml of protein solution with 0.001 ml of reservoir solution containing 0.1 M sodium chloride, 0.1 M trisodium citrate, pH 5.5, and 20% PEG 400, 18°C, to obtain a complex with cholest-4-en-3-one, a T104L mutant is used, and the crystals are grown in conditions containing 0.1 M MES, pH 6.0, and 18% PEG 550 monomethylether and soaked with 20% glycerol. For the T104L mutant-7KCh complex, crystals are grown in 0.1 M sodium chloride, 0.1 M tri-sodium citrate, pH 5.6, and 20% PEG 400, room temperature, X-ray diffraction structure determination and analysis at 1.90-2.75 A resolution