BRENDA - Enzyme Database show
show all sequences of 1.1.1.270

Solubilization and partial purification of a microsomal 3-ketosteroid reductase of cholesterol biosynthesis

Billheimer, J.T.; Alcorn, M.; Gaylor, J.L.; Arch. Biochem. Biophys. 211, 430-438 (1981)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
cytosolic Z protein
fatty acid-binding protein, membrane-bound enzyme, twofold to threefold activation, stimulatory activity is lost after solubilization of microsomal enzyme. Stimulation reversed by titration of Z-protein with either fatty acids or anti-Z-protein immunoglobulin. Stimulation not restored by incorporating partially purified reductase into an artificial phospholipid membrane
Rattus norvegicus
additional information
no activation by greatly increasing detergent concentration, Triton WR-1339, to a final concentration of 0.6%, no or poor activation by intestinal bile acid sequestrant, cholestyramine, no activation by bovine serum albumin
Rattus norvegicus
Application
Application
Commentary
Organism
analysis
enzyme can be used for reconstitution of 4-methyl sterol demethylations of cholesterol biosynthesis from lanosterol
Rattus norvegicus
General Stability
General Stability
Organism
enzyme extreme labile
Rattus norvegicus
enzyme stable in presence of high concentrations of detergents
Rattus norvegicus
ethylene glycol stabilizes
Rattus norvegicus
stable against subtilisin VII, no attendent loss of 3-ketoreductase activity
Rattus norvegicus
Inhibitors
Inhibitors
Commentary
Organism
Structure
ionic detergents
inactivation by strong solutions of ionic detergents, ethylene glycol minimizes inactivation
Rattus norvegicus
additional information
no inhibition by greatly increasing detergent concentration, Triton WR-1339, to a final concentration of 0.6%
Rattus norvegicus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.078
-
4alpha-methyl-5alpha-cholest-7-en-3-one
solubilized enzyme
Rattus norvegicus
0.236
-
4alpha-methyl-5alpha-cholest-7-en-3-one
microsomal-bound enzyme
Rattus norvegicus
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
membrane-bound
Rattus norvegicus
16020
-
microsome
microsomal-bound
Rattus norvegicus
-
-
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
chromatography on Sepharose 2B column shows ketoreductase activity eluted as high-molecular-weight complex
Rattus norvegicus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
additional information
Rattus norvegicus
catalyzes NADPH-dependent reduction of 3-ketosteroid intermediates, 4-methyl sterol intermediates, of cholesterol biosynthesis from lanosterol, regenerates 3beta-hydroxy sterol
?
-
-
-
additional information
Rattus norvegicus
enzyme of cholesterol biosynthesis
?
-
-
-
additional information
Rattus norvegicus
involved in the 10-step oxidative removal of the 4-gem-dimethyl group of sterols
?
-
-
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Rattus norvegicus
-
male sprague-dawley rats
-
Purification (Commentary)
Commentary
Organism
solubilization with Lubrol-WX in the presence of cholic acid, and partial purification
Rattus norvegicus
Source Tissue
Source Tissue
Commentary
Organism
Textmining
liver
-
Rattus norvegicus
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0014
-
partially purified enzyme
Rattus norvegicus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4,4-gem-dimethyl-5alpha-cholest-7-en-3-one + NADPH
-
286472
Rattus norvegicus
4,4-gem-dimethyl-5alpha-cholest-7-en-3beta-ol + NADP+
-
286472
Rattus norvegicus
?
4alpha-methyl-5alpha-cholest-7-en-3-one + NADPH
best substrate, reduction consistently about two times greater for monosubstituted steroid
286472
Rattus norvegicus
4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NADP+
-
286472
Rattus norvegicus
?
additional information
substrate specificity
286472
Rattus norvegicus
?
-
-
-
-
additional information
sterol substrate specificity with 4alpha-monomethyl- and 4,4-dimethyl-3-ketosteroids
286472
Rattus norvegicus
?
-
-
-
-
additional information
catalyzes NADPH-dependent reduction of 3-ketosteroid intermediates, 4-methyl sterol intermediates, of cholesterol biosynthesis from lanosterol, regenerates 3beta-hydroxy sterol
286472
Rattus norvegicus
?
-
-
-
-
additional information
enzyme of cholesterol biosynthesis
286472
Rattus norvegicus
?
-
-
-
-
additional information
involved in the 10-step oxidative removal of the 4-gem-dimethyl group of sterols
286472
Rattus norvegicus
?
-
-
-
-
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Rattus norvegicus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Rattus norvegicus
Cofactor
Cofactor
Commentary
Organism
Structure
additional information
essentially inactive with NADH
Rattus norvegicus
NADPH
specific for NADPH, NADH is essentially inactive, less than 10%
Rattus norvegicus
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
cytosolic Z protein
fatty acid-binding protein, membrane-bound enzyme, twofold to threefold activation, stimulatory activity is lost after solubilization of microsomal enzyme. Stimulation reversed by titration of Z-protein with either fatty acids or anti-Z-protein immunoglobulin. Stimulation not restored by incorporating partially purified reductase into an artificial phospholipid membrane
Rattus norvegicus
additional information
no activation by greatly increasing detergent concentration, Triton WR-1339, to a final concentration of 0.6%, no or poor activation by intestinal bile acid sequestrant, cholestyramine, no activation by bovine serum albumin
Rattus norvegicus
Application (protein specific)
Application
Commentary
Organism
analysis
enzyme can be used for reconstitution of 4-methyl sterol demethylations of cholesterol biosynthesis from lanosterol
Rattus norvegicus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
additional information
essentially inactive with NADH
Rattus norvegicus
NADPH
specific for NADPH, NADH is essentially inactive, less than 10%
Rattus norvegicus
General Stability (protein specific)
General Stability
Organism
enzyme extreme labile
Rattus norvegicus
enzyme stable in presence of high concentrations of detergents
Rattus norvegicus
ethylene glycol stabilizes
Rattus norvegicus
stable against subtilisin VII, no attendent loss of 3-ketoreductase activity
Rattus norvegicus
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
ionic detergents
inactivation by strong solutions of ionic detergents, ethylene glycol minimizes inactivation
Rattus norvegicus
additional information
no inhibition by greatly increasing detergent concentration, Triton WR-1339, to a final concentration of 0.6%
Rattus norvegicus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.078
-
4alpha-methyl-5alpha-cholest-7-en-3-one
solubilized enzyme
Rattus norvegicus
0.236
-
4alpha-methyl-5alpha-cholest-7-en-3-one
microsomal-bound enzyme
Rattus norvegicus
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
membrane-bound
Rattus norvegicus
16020
-
microsome
microsomal-bound
Rattus norvegicus
-
-
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
chromatography on Sepharose 2B column shows ketoreductase activity eluted as high-molecular-weight complex
Rattus norvegicus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
additional information
Rattus norvegicus
catalyzes NADPH-dependent reduction of 3-ketosteroid intermediates, 4-methyl sterol intermediates, of cholesterol biosynthesis from lanosterol, regenerates 3beta-hydroxy sterol
?
-
-
-
additional information
Rattus norvegicus
enzyme of cholesterol biosynthesis
?
-
-
-
additional information
Rattus norvegicus
involved in the 10-step oxidative removal of the 4-gem-dimethyl group of sterols
?
-
-
-
Purification (Commentary) (protein specific)
Commentary
Organism
solubilization with Lubrol-WX in the presence of cholic acid, and partial purification
Rattus norvegicus
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
liver
-
Rattus norvegicus
-
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0014
-
partially purified enzyme
Rattus norvegicus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4,4-gem-dimethyl-5alpha-cholest-7-en-3-one + NADPH
-
286472
Rattus norvegicus
4,4-gem-dimethyl-5alpha-cholest-7-en-3beta-ol + NADP+
-
286472
Rattus norvegicus
?
4alpha-methyl-5alpha-cholest-7-en-3-one + NADPH
best substrate, reduction consistently about two times greater for monosubstituted steroid
286472
Rattus norvegicus
4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NADP+
-
286472
Rattus norvegicus
?
additional information
substrate specificity
286472
Rattus norvegicus
?
-
-
-
-
additional information
sterol substrate specificity with 4alpha-monomethyl- and 4,4-dimethyl-3-ketosteroids
286472
Rattus norvegicus
?
-
-
-
-
additional information
catalyzes NADPH-dependent reduction of 3-ketosteroid intermediates, 4-methyl sterol intermediates, of cholesterol biosynthesis from lanosterol, regenerates 3beta-hydroxy sterol
286472
Rattus norvegicus
?
-
-
-
-
additional information
enzyme of cholesterol biosynthesis
286472
Rattus norvegicus
?
-
-
-
-
additional information
involved in the 10-step oxidative removal of the 4-gem-dimethyl group of sterols
286472
Rattus norvegicus
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Rattus norvegicus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Rattus norvegicus
Other publictions for EC 1.1.1.270
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
740936
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4-Methylzymosterone and other ...
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741250
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e113194
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726098
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Role of sterol 3-ketoreductase ...
Botrytis cinerea
Pest Manag. Sci.
69
642-651
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740097
Layer
Characterization of a mutation ...
Mus musculus
Biochim. Biophys. Acta
1831
361-369
2013
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724124
Endo
Characterization of rabbit ald ...
Oryctolagus cuniculus
Arch. Biochem. Biophys.
527
23-30
2012
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1
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4
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14
20
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27
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14
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6
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27
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1
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3
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702462
Taramino
Interactions of oxidosqualene ...
Saccharomyces cerevisiae
Biochim. Biophys. Acta
1801
156-162
2010
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711300
Taramino
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Homo sapiens, Mus musculus
Biochim. Biophys. Acta
1801
1232-1237
2010
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2
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7
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1
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725813
Thomas
The functions of key residues ...
Homo sapiens
J. Steroid Biochem. Mol. Biol.
120
192-199
2010
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1
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6
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3
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673822
Tam
Activities of 3beta-HSD and ar ...
Taeniopygia guttata
Gen. Comp. Endocrinol.
150
26-33
2006
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675595
Steckelbroeck
Tibolone metabolism in human l ...
Homo sapiens
J. Pharmacol. Exp. Ther.
316
1300-1309
2006
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677144
Jin
Molecular docking simulations ...
Homo sapiens
Steroids
71
380-391
2006
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656200
Steckelbroeck
Human cytosolic 3alpha-hydroxy ...
Homo sapiens
J. Biol. Chem.
279
10784-10795
2004
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655348
Rizner
Human type 3 3alpha-hydroxyste ...
Homo sapiens
Endocrinology
144
2922-2932
2003
-
-
-
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1
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656795
Marijanovic
Closing the gap: Identificatio ...
Homo sapiens, Mus musculus
Mol. Endocrinol.
17
1715-1725
2003
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2
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1
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5
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723672
Mo
Protein-protein interactions a ...
Saccharomyces cerevisiae
Proc. Natl. Acad. Sci. USA
99
39-44
2002
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