Information on EC 1.14.15.6 - cholesterol monooxygenase (side-chain-cleaving)

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The expected taxonomic range for this enzyme is: Coelomata

EC NUMBER
COMMENTARY hide
1.14.15.6
-
RECOMMENDED NAME
GeneOntology No.
cholesterol monooxygenase (side-chain-cleaving)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(20R,22R)-20,22-dihydroxy-cholesterol + 2 reduced adrenodoxin + O2 + 2 H+ = pregnenolone + 4-methylpentanal + 2 oxidized adrenodoxin + 2 H2O
show the reaction diagram
(1c)
-
-
-
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2 + 2 H+ = (20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
(1b)
-
-
-
cholesterol + 2 reduced adrenodoxin + O2 + 2 H+ = (22R)-22-hydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
(1a)
-
-
-
cholesterol + 6 reduced adrenodoxin + 3 O2 + 6 H+ = pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O
show the reaction diagram
overall reaction
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-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
androgen and estrogen metabolism
-
-
Metabolic pathways
-
-
Steroid hormone biosynthesis
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-
SYSTEMATIC NAME
IUBMB Comments
cholesterol,reduced-adrenal-ferredoxin:oxygen oxidoreductase (side-chain-cleaving)
A heme-thiolate protein (cytochrome P-450). The reaction proceeds in three stages, with two hydroxylations at C-22 and C-20 preceding scission of the side-chain between carbons 20 and 22. The initial source of the electrons is NADPH, which transfers the electrons to the adrenodoxin via EC 1.18.1.6, adrenodoxin-NADP+ reductase.
CAS REGISTRY NUMBER
COMMENTARY hide
37292-81-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
yellowtail clownfish, an protandrous hemaphroditic, anemone-symbiontic fish
-
-
Manually annotated by BRENDA team
Japanese eel, gene CYP11A1
SwissProt
Manually annotated by BRENDA team
hircus
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
non-breeding male
-
-
Manually annotated by BRENDA team
-
Swissprot
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
synthetic construct
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
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
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(20R,22R)-20,22-dihydroxy-cholesterol + 2 reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + 2 oxidized adrenodoxin + 2 H2O
show the reaction diagram
(20S)-22-thiacholesterol + reduced adrenodoxin + O2
(20S,22R)-22-thiacholesterol S-oxide + (20S,22S)-22-thiacholesterol S-oxide
show the reaction diagram
-
-
(22R)-sulfoxide preferentially formed by a factor of 4.2 to 1 over (22S)-sulfoxide
?
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2
(20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
17,20-dihydroxyvitamin D2 + reduced adrenodoxin + O2
17,20,24-trihydroxyvitamin D2 + oxidized adrenodoxin + O2
show the reaction diagram
-
is slowly metabolized
-
-
?
20,23-dihydroxyvitamin D3 + reduced adrenodoxin + O2
17,20,23-trihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20,23-dihydroxyvitamin D3 + reduced adrenodoxin + O2
17alpha,20,23-trihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20-hydroxyvitamin D2 + reduced adrenodoxin + O2
17,20,24-trihydroxyvitamin D2 + oxidized adrenodoxin + O2
show the reaction diagram
-
is a better substrate than vitamin D2 itself
-
-
?
20-hydroxyvitamin D3 + reduced adrenodoxin + O2
20,23-dihydroxyvitamin D3 + 17alpha,20,23-trihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20-hydroxyvitamin D3 + reduced adrenodoxin + O2
20,23-dihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20alpha, 22(R)-dihydroxycholesterol + reduced adrenodoxin + O2
? + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20alpha-hydroxycholesterol + reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20alpha-hydroxycholesterol + reduced adrenodoxin + O2
pregnenolone + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
?
22(R)-hydroxycholesterol + reduced adrenodoxin + O2
? + oxidized adrenodoxin + H2O
show the reaction diagram
22(R)-hydroxycholesterol + reduced adrenodoxin + O2
pregnenolone + oxidized adrenodoxin + H2O
show the reaction diagram
25-hydroxycholesterol + O2 + reduced ferredoxin
pregnenolone + H2O + ferredoxin + 4-hydroxy-4-methylpentanal
show the reaction diagram
-
-
-
-
?
25-hydroxycholesterol + reduced adrenodoxin + O2
pregnenolone + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
?
cholesterol + 2 reduced adrenodoxin + O2
(22R)-22-hydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
cholesterol + 6 reduced adrenodoxin + 3 O2
pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O
show the reaction diagram
cholesterol + 6 reduced adrenodoxin mutant S112W + 3 O2
pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin mutant S112W + 4 H2O
show the reaction diagram
-
overall reaction, adrenodoxin mutant S112W gives a 14fold higher catalytic efficiency compared to wild-type adrenodoxin
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-
?
cholesterol + reduced adrenal ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenal ferredoxin
show the reaction diagram
cholesterol + reduced adrenal ferredoxin + O2
pregnenolone + adrenal ferredoxin + H2O
show the reaction diagram
cholesterol + reduced adrenodoxin + O2
? + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
-
cholesterol + reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenodoxin + H2O
show the reaction diagram
cholesterol + reduced ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized ferredoxin
show the reaction diagram
cholesterol sulfate + reduced adrenodoxin + O2
pregnenolone sulfate + 17-hydroxy-pregnenolone + dehydroisoandrosterone sulfate + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
?
vitamin D2 + reduced adrenodoxin + O2
17,20,24-trihydroxyvitamin D2 + oxidized adrenodoxin + H2O
show the reaction diagram
-
P450scc catalyzes three sequential hydroxylations of D2 producing 20-hydroxyvitamin D2, 17,20-dihydroxyvitamin D2, and 17,20,24-trihydroxyvitamin D2, which dissociate from the active site of P450scc and accumulate in the reaction mixture
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-
?
vitamin D3 + reduced adrenodoxin + O2
20-hydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(20R,22R)-20,22-dihydroxy-cholesterol + 2 reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + 2 oxidized adrenodoxin + 2 H2O
show the reaction diagram
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2
(20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
cholesterol + 2 reduced adrenodoxin + O2
(22R)-22-hydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
cholesterol + 6 reduced adrenodoxin + 3 O2
pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O
show the reaction diagram
cholesterol + reduced adrenal ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenal ferredoxin
show the reaction diagram
-
first and rate-limiting enzyme in adrenal steroidogenesis
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-
?
cholesterol + reduced adrenal ferredoxin + O2
pregnenolone + adrenal ferredoxin + H2O
show the reaction diagram
cholesterol + reduced ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized ferredoxin
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adrenodoxin
cytochrome P450
Ferredoxin
Iron
-
the enzyme is a cytochrome P450 enzyme
reduced adrenal ferredoxin
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(20R,S)-20-amino-5-pregnen-3beta-ol
-
20-amine derivative, amine is attached closer to the D-ring than in the 22-amine, very weak inhibitor, 0.1 mM causes less than 20% inhibition
-
(20S)-22-thiacholesterol
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competitive inhibitor, is converted enzymatically to a more potent inhibitor, the (22S) and (22R) sulfoxides, inhibition by approximately 75% at 0.001 mM, no inactivation in absence of NADPH and O2
(20S,22R)-22-thiacholesterol S-oxide
-
competitive versus cholesterol, binds 10times more tightly than (22S) diastereomer, 25% and 44% inhibition at 0.00005 mM and 0.0001 mM, respectively, complete inhibition at 0.001 mM, no substrate for P-450
(20S,22S)-22-thiacholesterol S-oxide
-
competitive versus cholesterol, no substrate for P-450
(22R)-22-Aminocholesterol
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completely inhibited by 0.001 mM, affinity toward the P-450scc almost 3fold greater than that for the (22S) form, competitive versus cholesterol, no substrate for P-450
(22S)-22-Aminocholesterol
-
not inhibited below 0.001 mM, competitive versus cholesterol, no substrate for P-450
17,20-dihydroxyvitamin D2
-
competitive inhibitor
17beta-amino-5-androsten-3beta-ol
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17-amine derivative, amine is attached closer to the D-ring than in the 22-amine, very weak inhibitor, 0.1 mM causes less than 20% inhibition
22-Amino-23,24-bisnor-5-cholen-3beta-ol
22-amino-23,24-bisnor-5alpha-cholen-3beta-ol
-
50% inhibition at 0.003 mM
23,24-bisnor-5-cholene-3beta,22-diol
-
competitive inhibitor, 40% inhibition at 0.01 mM, 50% at 0.015 mM, resembles the intermediate 22-hydroxycholesterol but acts as an inhibitor rather than serving as a substrate
23-Amino-24-nor-5-cholen-3beta-ol
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23-amine derivative, same steroid ring structure as cholesterol, competitive inhibitor with respect to cholesterol, 50% inhibition at 0.0001 mM, reversible cooperative binding
24-Amino-5-cholen-3beta-ol
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24-amine derivative, amine attached in greater distance from steroid ring, same steroid ring structure as cholesterol, causes a progressive decrease in inhibitory potency, 50% inhibition at 0.0023 mM, reversible noncooperative binding
25-Amino-26,27-bisnor-5-cholesten-3beta-ol
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25-amine derivative, amine attached in greater distance from steroid ring, causes a progressive decrease in inhibitory potency, 50% inhibition at more than 0.1 mM
adrenodoxin
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oxidized form, high affinity to P-450scc, inhibits side chain cleavage by competition with reduced form
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aminogluthetimide
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-
cholesterol
-
inhibition above 0.003 mM, mitochondrial
Cholesterol sulfate
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inhibition above 0.005 mM, mitochondrial
glycerol
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substrate cholesterol, 50% inhibition at 25% glycerol, substrate 22(R)-hydroxycholesterol, 50% inhibition at 44% glycerol, substrate 20alpha-hydroxycholesterol, 50% inhibition at 48% glycerol, substrate 20alpha, 22(R)-dihydroxycholesterol, 50% inhibition at 51% glycerol
methoxychlor
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pesticide of DDT, suicide inhibitor, competitive to cholesterol, substantial irreversible loss of activity, 5% inhibition within 5 min at 0.2 mM, decrease is suppressed by the presence of cholesterol
phosphatidyl choline
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-
phosphatidyl ethanolamine
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-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,2-di-(2'-hexyl-decanoyl)-sn-glycero-3-phosphocholine
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alpha-branched phosphatidylcholine, inclusion in vesicle-reconstituted system, partially in connection with the nonactivator lipids dimyristoyl-/dioleoyl-phosphatidylcholine, efficiency close to cardiolipin
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1,2-di-(2'-octyl-dodecanoyl)-sn-glycero-phosphocholine
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alpha-branched phosphatidylcholine, inclusion in vesicle-reconstituted system, partially in connection with the nonactivator lipids dimyristoyl-/dioleoyl-phosphatidylcholine, efficiency close to cardiolipin
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Brij
-
56, 76 and 96
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cardiolipin
Emulgen
-
911 and 913
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fatty acid
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C18, natural detergents in DMPC vesicles, stimulation similar to octyl glucoside
glycerol
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5-20% concentration, 20-50% increase of enzyme activity
Lipid
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from adrenal mitochondria accelerates activity
octyl glucoside
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high concentrations of this detergent cause 50% stimulation of cholesterol side-chain-cleavage in large unilamellar vesicles at low cholesterol concentration, 0.01 mM increase the proportion of P-450 bound by cholesterol
Phospholipid
Triton X-100
-
effect onto P-450 itself suggested
Tween 20
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0181
17,20-dihydroxyvitamin D2
-
in 0.45% cyclodextrin
0.012
20-hydroxyvitamin D2
-
in 0.45% cyclodextrin
0.067
20-hydroxyvitamin D3
-
in the presence of 0.45% cyclodextrin
0.012
20alpha-hydroxycholesterol
-
-
0.0004 - 0.0012
adrenodoxin
0.0005 - 1.18
cholesterol
0.0003 - 0.0232
Cholesterol sulfate
0.0175
vitamin D2
-
in 0.45% cyclodextrin
0.0296 - 3.67
vitamin D3
additional information
additional information
-
decrease in concentration of adrenodoxin reductase causes a decrease in Km-values for cholesterol and adrenodoxin, in mitochondria, cholesterol is near-saturating for enzyme activity due to low and rate-limiting concentration of adrenodoxin reductase present
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.123
20-hydroxyvitamin D3
Bos taurus
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in the presence of 0.45% cyclodextrin
0.05 - 0.5
cholesterol
0.328 - 1.6
vitamin D3
additional information
additional information
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.83
20-hydroxyvitamin D3
Bos taurus
-
in the presence of 0.45% cyclodextrin
10258
0.212 - 11.17
cholesterol
0.435 - 11.1
vitamin D3
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8
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for cholesterol and 20alpha-hydroxycholesterol, 75 mM potassium phosphate buffer
7.3
-
assay at
7.4
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7.5
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less than 50% of maximal activity below and above range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 42
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tested in different dimyristoyl-phosphatidylcholine vesicles, breaks in activity at 27-30C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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low level
Manually annotated by BRENDA team
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epithelial ovarian carcinoma
Manually annotated by BRENDA team
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basophilic cells, exclusively
Manually annotated by BRENDA team
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pre-ovulatory follicle F1
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
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breast cancer cell, expresses both the 55-kDa isoform and the 32-kDa truncated isoform of CYP11A1
Manually annotated by BRENDA team
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adrenocortical cell line
Manually annotated by BRENDA team
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dorsal root ganglion, spinal cord, dorsal horn, nociceptive supraspinal nuclei and somatosensory cortex
Manually annotated by BRENDA team
-
granulosa cell
Manually annotated by BRENDA team
-
expresses the full length 55-kDa isoform of CYP11A1
Manually annotated by BRENDA team
-
low level
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
truncated form
Manually annotated by BRENDA team
additional information
synthetic construct
-
all of the addressing sequences, containing transmembrane domains, provide effective insertion of the hybrid proteins AAC-mCYP11A1, Bcs1p(1-83)-mCYP11A1, DLD(1-72)-mCYP11A1 and Su9(1-116)-mCYP11A1 into the mitochondrial inner membrane. preAd-mCYP11A1 hybrid molecules are translocated across the inner membrane and tightly associated with the membrane on its matrix side but not membrane inserted. The mechanism of Ad-mCYP11A1 hybrid topogenesis in Escherichia coli cells differs from that of the topogenesis of its precursor form in yeast mitochondria
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
97000
-
sedimentation equilibrium
100000
-
native-PAGE, incomplete enzyme or molecular environmental conditions not optimal
200000
220000
-
corpus luteum
225000
-
sedimentation equilibrium
400000
-
native-PAGE
415000
-
sedimentation equilibrium
470000
-
sedimentation equilibrium performed in 100 mM potassium phosphate buffer, pH 7.6
800000
-
exclusion chromatography
850000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexadecamer
-
16 * 53000, sedimentation equilibrium performed in 6 M guanidine after heating, 16 * 52000, SDS-PAGE. Can also exist in forms of 4 and 8 subunits after treatment with 100 mM potassium phosphate buffer, 7.6, with molecular weights of 200000 and 470000, respectively
tetramer
-
4 * 46000, sedimentation equilibrium analysis after guanidine treatment and SDS-PAGE
additional information
-
the enzyme structure possesses a substrate access channel, the F-G loop is a membrane-interacting area
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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, 18C, overnight, X-ray diffraction structure determination and analysis at 2.8 A resolution, molecular replacement using rat mitochondrial CYP24A1
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
increase of complex formation of adrenodoxin and cytochrome P-450scc in absence of cholesterol with decrease of temperature to 6C, elevated temperature decreases the affinity of P-450scc for cholesterol
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
catalase, protects P-450scc against activity loss
-
glycerol, 50%, stabilizes
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Glycerol
-
induces sigmoidal low-spin response of otherwise high-spin spectrum, weakens adrenodoxin binding, inhibitory effect being substrate dependent
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 10 mM sodium phosphate buffer, pH 7.4, 0.1 mM EDTA, 20% glycerol, stable for months
-
-20C, 50 mM potassium phosphate buffer, pH 7.0, 50% glycerol, 3 months
-
-20C, 50% glycerol, for months
-
-20C, mitochondrial precipitate, 10 mM sodium phosphate buffer, pH 7.4, 0.1 mM EDTA
-
-20C, purified, 10 mM sodium phosphate buffer, pH 7.4, 0.1 mM EDTA, 20% glycerol, for months without loss of P-450 activity
-
-70C, mitochondrial pellet, 30-40 mg/ml, 100 mM potassium phosphate, pH 7.3, 0.2 mM EDTA
-
-80C, 50 mM potassium phosphate buffer, pH 7.0, 0.1 mM EDTA, 0.1 mM dithiothreitol, 0.01% cholate, a few days without substantial loss of activity
-
0C on ice, purified, 20 mM potassium phosphate buffer, pH 7.4, 20% glycerol, v/v, 0.1 mM EDTA, 0.1 mM dithioerythritol, 0.01% Emulgen 911, 3 weeks, without substantial loss of activity
-
4C, purified, 50 mM potassium phosphate, pH 7.3, 0.1 mM dithiothreitol, 10% glycerol, stable a few weeks
-
5C, 50 mM potassium phosphate buffer, pH 7.0, without glycerol, 21-43% decrease of activity within 30 days, completely lost in 4 months
-
purified, 25 mM potassium phosphate, pH 7.3, 0.05 mM EDTA, 0.05 mM dithiothreitol, 0.025 mM deoxycorticosterone, 0.25% Tween 20, 0.25% sodium cholate
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography
-
ammonium sulfate fractioning, aniline-Sepharose, adrenodoxin-Separose
-
ammonium sulfate precipitation, affinity chromatography
-
ammonium sulfate precipitation, affinity chromatography; DEAE-cellulose, adrenodoxin-Sepharose
-
cholate extraction, pyrophosphate treatment, affinity chromatography
-
cholic acid extraction, ammonium sulfate precipitation, DEAE-cellulose, hydroxylapatite and gel filtration
-
column chromatography, gel filtration
-
DEAE-cellulose, adrenodoxin-Sepharose
-
iso-octane and ammonium sulfate fractionating, gel filtration
-
native enzyme partially by purification of mitochondrial inner membranes
-
precipitation with polyethylene glycol, affinity chromatography
-
recombinant C-terminally His4-tagged CYP11A1 by anion exchange, nickel affinity, hydroxylapatite, adrenodoxin affinity chromatography, and gel filtration
recombinant His-tagged CYP11A1 protein from Escherichia coli by nickel affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
several ion exchange and heptyl-Sepharose hydrophobic chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
both full-length and an N-terminally truncated form amplified and inserted into TOPO TA PCR cloning vector and subsequently subcloned into the EcoRI restriction site of pCMV-tag2B vector to obtain CYP11A1 proteins with an N-terminal Flag tag or into pEGFP-C2 vector to yield an N-terminal GFP tag
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DNA and amino acid sequence determination and analysis, expression analysis and sequence comparisons
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DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis, and expression in COS-1 cells with adrenodoxin resulting in significantly more pregnenolone, co-expression with the steroidogenic acute regulatory protein and a human P450scc/adrenodoxin reductase/adrenodoxin fusion constructs. COS-1 cells transfected with a modified construct in which human P450scc is replaced by Dasyatis sabina P450scc show higher rates than cells transfected with Dasyatis sabina P450scc alone. Suncloning in Escherichia coli strain DH5alpha
DNA sequence determination, quantitative enzyme expression analysis in brain prior and after treatment with 4-nonylphenol
-
expressed from pCMV-SPORT6 vector
-
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene CYP11A1, DNA and amino acid sequence determination and analysis, expression pattern analysis, functional expression in COS-7 cells
-
gene CYP11A1, DNA and amino acid sequence determination of wild-type and mutant enzymes, expression in HeLa cells and COS-1 cells, functional studies of P450scc RNA splicing
gene CYP11A1, expression as His-tagged protein in Escherichia coli
-
gene p450scc, semi-/quantitative RT-PCR expression analysis, overview
-
heterologous expression in Escherichia coli
-
hybrid proteins AAC-mCYP11A1, Bcs1p(183)-mCYP11A1, Su9(1-116)-mCYP11A1, preAd-mCYP11A1, DLD(1-72)-mCYP11A1 and COXIV-mCYP11A1 expressed in Saccharomyces cerevisiae strain 2805. Escherichia coli strain JM109 transformed with the plasmid pTrc99(A)/Ad-mCYP11A1
synthetic construct
-
mutants and wild-type expressed in Escherichia coli
-
Nicotiana tabacum plants expressing CYP11A1 cDNA
-
recombinant expression of C-terminally His4-tagged CYP11A1
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
14 days exposure of previtellogenic ovarian oocytes to nonylphenol produces a unique and consistent concentration-specific pattern of modulation of StAR and P450scc gene expression, increasing from 0 (control) to 1 and 10 micromol, and decreasing at 50 and 100 micromol
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contents of soluble protein and carbohydrates in leaves and seeds of transgenic Nicotiana tabacum plants are essentially higher than the contents of these components in leaves and seeds of control plants
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CYP11A1 expression is induced by Runx2in osteoblasts
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CYP11A1 is elevated in dominant follicles. CYP11A1 mRNA levels are highly correlated with the focimatrix genes COL4A1, NID1 and -2 and HSPG2. Focimatrix may potentially regulate CYP11A1 expression, and the regulation of both may be important in follicular dominance
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CYP11A1 mRNA expression is dramatically induced within 2 h after introduction of Runx2 protein into Runx2-null cells. CYP11A1 gene expression is induced during osteoblastic differentiation
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expression of StAR and P450scc transcripts in non-steroidogenic tissues
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induction of P450scc mRNA expression by 8-Br-cAMP
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mRNA levels for CYP11A1 are significantly lower in subordinate follicles in comparison to dominant follicles and this effect is maintained following adjustment for follicle size
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P450scc expression is reduced in Leydig cells treated with synthetic hormone diethylstilbestrol and natural hormone 17beta-estradiol. Diethylstilbestrol causes histone deacetylation in the P450scc promoter region, while 17beta-estradiol does not
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relationship between cortisol production and induction of transcription of steroidogenic acute regulatory (StAR) protein and P450scc. Exposure of juveniles to 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene and the synthetic estrogen 17alpha-ethynylestradiol produces transcriptional modulations of StAR and P450scc expression in interrenal tissue. In juvenile salmon exposed to 17alpha-ethynylestradiol, expression of P450scc mRNA is detectable in the trunk kidney, while StAR mRNA is not quantifiable. 17alpha-ethynylestradiol produces time- and concentration-specific effects on the expression of the StAR, P450scc, P450arom isoforms and IGF-2 genes in gonadal tissues. In liver and kidney tissues exposed to the endocrine disrupting chemical 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene, significant induction of both StAR and P450scc mRNA
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testosterone significantly increases P450scc in F1 granulosa cells after 3 h at 10 and 100 ng/ml
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K267Q
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participation in electrostatic interaction of P-450scc with adrenodoxin
K338Q
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removed from heme group but still very important for interaction with adrenodoxin, K helix
K342Q
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removed from heme group but still very important for interaction with adrenodoxin, K helix
R425Q
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most harmful substitution, L helix, heme-binding region
R425Q/R426C
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double mutant, most harmful substitution
R425Q/R426Q
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double mutant, most harmful substitution
c835delA
a naturally occuring CYP11A1 frameshift mutation, heterozygous mutant, phenotype, overview
L141W
a naturally occuring CYP11A1 missense mutation, heterozygous mutant, phenotype, overview
V415E
a naturally occuring CYP11A1 missense mutation, heterozygous mutant, phenotype, overview
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
synthetic construct
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the development of a cholesterol biosensor based on screen-printed electrodes modified with multi-walled carbon nanotubes and with the cytochromes P450scc may ensure a high sensitivity. Role of the nanotubes in mediating electron transfer to the cytochrome P450scc is verified as further improved with respect to the case of rhodium-graphite electrodes modified by the use of gold nanoparticles
drug development
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the vitamin D3 derivatives produced by the action of P450scc are good candidates for use in the therapy of hyperproliferative disorders
medicine