Literature summary for 1.14.15.6 extracted from

Mast, N.; Annalora, A.; Lodowski, D.; Palczewski, K.; Stout, C.; Pikuleva, I.
Structural basis for three-step sequential catalysis by the cholesterol side chain cleavage enzyme CYP11A1 (2011), J. Biol. Chem., 286, 5607-5613.

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Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of C-terminally His4-tagged CYP11A1	Bos taurus

Crystallization (Commentary)

Crystallization (Comment)	Organism
CYP11A1 in complex with (22R)-22-hydroxycholesterol, sitting drop vapour diffusion methdo, mixing of 0.001 ml of 23 mg/ml protein in 50 mM potassium phosphate, pH 7.2, 20% glycerol, 0.1 M NaCl, 0.1% octyl pentaethylene glycol ether, 1 mM EDTA, and 0.05 mM (22R)-22-hydroxycholesterol, with 0.001 ml of precipitant solution containing 14% PEG 1000, 20% glycerol, 12% JEFFAMINE ED-2001, 0.1 M MES, pH 7.0, and 10% isopropyl alcohol, 18°C, overnight, X-ray diffraction structure determination and analysis at 2.8 A resolution, molecular replacement using rat mitochondrial CYP24A1	Bos taurus

Crystallization (Comment)

Organism

CYP11A1 in complex with (22R)-22-hydroxycholesterol, sitting drop vapour diffusion methdo, mixing of 0.001 ml of 23 mg/ml protein in 50 mM potassium phosphate, pH 7.2, 20% glycerol, 0.1 M NaCl, 0.1% octyl pentaethylene glycol ether, 1 mM EDTA, and 0.05 mM (22R)-22-hydroxycholesterol, with 0.001 ml of precipitant solution containing 14% PEG 1000, 20% glycerol, 12% JEFFAMINE ED-2001, 0.1 M MES, pH 7.0, and 10% isopropyl alcohol, 18°C, overnight, X-ray diffraction structure determination and analysis at 2.8 A resolution, molecular replacement using rat mitochondrial CYP24A1

Bos taurus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
mitochondrion	-	Bos taurus	5739	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
(20R,22R)-20,22-dihydroxy-cholesterol + 2 reduced adrenodoxin + O2	Bos taurus	-	pregnenolone + 4-methylpentanal + 2 oxidized adrenodoxin + 2 H2O	-	?
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2	Bos taurus	-	(20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O	-	?
cholesterol + 2 reduced adrenodoxin + O2	Bos taurus	-	(22R)-22-hydroxycholesterol + 2 oxidized adrenodoxin + H2O	-	?
cholesterol + 6 reduced adrenodoxin + 3 O2	Bos taurus	overall reaction	pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Bos taurus	P00189	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant C-terminally His4-tagged CYP11A1 by anion exchange, nickel affinity, hydroxylapatite, adrenodoxin affinity chromatography, and gel filtration	Bos taurus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(20R,22R)-20,22-dihydroxy-cholesterol + 2 reduced adrenodoxin + O2	-	Bos taurus	pregnenolone + 4-methylpentanal + 2 oxidized adrenodoxin + 2 H2O	-	?
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2	-	Bos taurus	(20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O	-	?
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2	product binding structure analysis, detailed overview	Bos taurus	(20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O	-	?
cholesterol + 2 reduced adrenodoxin + O2	-	Bos taurus	(22R)-22-hydroxycholesterol + 2 oxidized adrenodoxin + H2O	-	?
cholesterol + 6 reduced adrenodoxin + 3 O2	overall reaction	Bos taurus	pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O	-	?

Synonyms

Synonyms	Comment	Organism
cholesterol side chain cleavage enzyme	-	Bos taurus
CYP11A1	-	Bos taurus
cytochrome P450 11A1	-	Bos taurus
P450 11A1	-	Bos taurus

Cofactor

Cofactor	Comment	Organism
adrenodoxin	-	Bos taurus
cytochrome P450	-	Bos taurus
heme	site of catalysis	Bos taurus

General Information

General Information	Comment	Organism
additional information	the active site cavity in CYP11A1 represents a long curved tube that extends from the protein surface to the heme group, the site of catalysis. Shuttling of the sterol intermediates between the active site entrance and the heme group during the three-step reaction. Structural basis of the strict substrate specificity and high catalytic efficiency of the enzyme, overview	Bos taurus