BRENDA - Enzyme Database
show all sequences of 1.14.15.28

Structural and biochemical characterization of Mycobacterium tuberculosis CYP142: evidence for multiple cholesterol 27-hydroxylase activities in a human pathogen

Driscoll, M.D.; McLean, K.J.; Levy, C.; Mast, N.; Pikuleva, I.A.; Lafite, P.; Rigby, S.E.; Leys, D.; Munro, A.W.; J. Biol. Chem. 285, 38270-38282 (2010)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
-
Mycobacterium tuberculosis
Crystallization (Commentary)
Crystallization
Organism
sitting drop method, the CYP142 crystal structure is solved to 1.6 A
Mycobacterium tuberculosis
General Stability
General Stability
Organism
completely to the P450 state on binding of cholest-4-en-3-one at pH 8.0
Mycobacterium tuberculosis
stabilizing effect of substrate binding on the thiolate-coordinated CYP142, to the extent that the P420 form of CYP142 can be converted almost
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
Mycobacterium tuberculosis
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
Mycobacterium tuberculosis ATCC 25618
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mycobacterium tuberculosis
P9WPL5
-
-
Mycobacterium tuberculosis ATCC 25618
P9WPL5
-
-
Purification (Commentary)
Commentary
Organism
-
Mycobacterium tuberculosis
Reaction
Reaction
Commentary
Organism
(25R)-26-hydroxycholest-4-en-3-one + 2 reduced [2Fe-2S] ferredoxin + O2 = (25R)-26-oxocholest-4-en-3-one + 2 oxidized [2Fe-2S] ferredoxin + 2 H2O
(1b)
Mycobacterium tuberculosis
(25R)-26-oxocholest-4-en-3-one + 2 reduced [2Fe-2S] ferredoxin + O2 = (25R)-3-oxocholest-4-en-26-oate + 2 oxidized [2Fe-2S] ferredoxin + H2O
(1c)
Mycobacterium tuberculosis
cholest-4-en-3-one + 2 reduced [2Fe-2S] ferredoxin + O2 = (25R)-26-hydroxycholest-4-en-3-one + 2 oxidized [2Fe-2S] ferredoxin + H2O
(1a)
Mycobacterium tuberculosis
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2 = (25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
overall reaction
Mycobacterium tuberculosis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
736407
Mycobacterium tuberculosis
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
catalyses the hydroxylation of the C-26 carbon, followed by oxidation of the alcohol to the carboxylic acid via the aldehyde intermediate, initiating the degradation of the alkyl side-chain of cholesterol. The products are exclusively in the (25R) conformation
736407
Mycobacterium tuberculosis
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
736407
Mycobacterium tuberculosis ATCC 25618
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
catalyses the hydroxylation of the C-26 carbon, followed by oxidation of the alcohol to the carboxylic acid via the aldehyde intermediate, initiating the degradation of the alkyl side-chain of cholesterol. The products are exclusively in the (25R) conformation
736407
Mycobacterium tuberculosis ATCC 25618
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholesterol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
736407
Mycobacterium tuberculosis
3beta-hydroxycholest-5-en-26-oic acid + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholesterol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
736407
Mycobacterium tuberculosis ATCC 25618
3beta-hydroxycholest-5-en-26-oic acid + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
6
-
the substrate free enzyme is unstable and aggregates
Mycobacterium tuberculosis
7
-
the P450 form of CYP142 is most stable at pH 7, and larger proportions of the P420 species are formed at the higher pH values, with near-complete P420 formation at pH 9
Mycobacterium tuberculosis
8
-
the spectrum for the Fe2+-CO form is notably unstable, and the P450 species progressively collapses over time with P420 accumulation
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
sitting drop method, the CYP142 crystal structure is solved to 1.6 A
Mycobacterium tuberculosis
General Stability (protein specific)
General Stability
Organism
completely to the P450 state on binding of cholest-4-en-3-one at pH 8.0
Mycobacterium tuberculosis
stabilizing effect of substrate binding on the thiolate-coordinated CYP142, to the extent that the P420 form of CYP142 can be converted almost
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
Mycobacterium tuberculosis
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
Mycobacterium tuberculosis ATCC 25618
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
736407
Mycobacterium tuberculosis
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
catalyses the hydroxylation of the C-26 carbon, followed by oxidation of the alcohol to the carboxylic acid via the aldehyde intermediate, initiating the degradation of the alkyl side-chain of cholesterol. The products are exclusively in the (25R) conformation
736407
Mycobacterium tuberculosis
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
the enzyme can receive electrons from ferredoxin reductase in vitro, its natural electron donor is not known yet. The enzyme of the bacterial pathogen is involved degradation of the host cholesterol
736407
Mycobacterium tuberculosis ATCC 25618
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced [2Fe-2S] ferredoxin + 3 O2
catalyses the hydroxylation of the C-26 carbon, followed by oxidation of the alcohol to the carboxylic acid via the aldehyde intermediate, initiating the degradation of the alkyl side-chain of cholesterol. The products are exclusively in the (25R) conformation
736407
Mycobacterium tuberculosis ATCC 25618
(25R)-3-oxocholest-4-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholesterol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
736407
Mycobacterium tuberculosis
3beta-hydroxycholest-5-en-26-oic acid + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholesterol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
736407
Mycobacterium tuberculosis ATCC 25618
3beta-hydroxycholest-5-en-26-oic acid + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
6
-
the substrate free enzyme is unstable and aggregates
Mycobacterium tuberculosis
7
-
the P450 form of CYP142 is most stable at pH 7, and larger proportions of the P420 species are formed at the higher pH values, with near-complete P420 formation at pH 9
Mycobacterium tuberculosis
8
-
the spectrum for the Fe2+-CO form is notably unstable, and the P450 species progressively collapses over time with P420 accumulation
Mycobacterium tuberculosis
General Information
General Information
Commentary
Organism
metabolism
the enzyme is involved in host response modulation and cholesterol metabolism
Mycobacterium tuberculosis
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the enzyme is involved in host response modulation and cholesterol metabolism
Mycobacterium tuberculosis
Other publictions for EC 1.14.15.28
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)
738629
Frank
Cholesterol ester oxidation by ...
Mycolicibacterium smegmatis
J. Biol. Chem.
289
30417-30425
2014
-
-
-
1
-
-
-
1
-
-
-
1
-
2
-
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1
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1
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1
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1
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1
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1
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1
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-
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-
738114
Garcia-Fernandez
A highly conserved mycobacteri ...
Mycolicibacterium smegmatis, Mycolicibacterium smegmatis mc(2)155
Environ. Microbiol.
15
2342-2359
2013
-
-
1
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-
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1
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2
-
11
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1
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2
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1
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1
1
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1
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2
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1
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2
-
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-
744481
Johnston
Substrate analog studies of t ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Bioorg. Med. Chem.
20
4064-4081
2012
-
-
-
-
-
-
-
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-
-
-
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3
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10
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4
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10
-
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4
-
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-
-
-
-
-
-
-
717811
Johnston
Functional redundancy of stero ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
285
36352-36360
2010
-
-
1
-
-
-
-
2
-
-
-
6
-
161
-
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1
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-
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12
-
1
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2
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1
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1
1
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2
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6
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1
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-
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12
-
1
-
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2
-
-
-
-
-
-
-
-
2
2
736407
Driscoll
Structural and biochemical cha ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis ATCC 25618
J. Biol. Chem.
285
38270-38282
2010
-
-
1
1
-
2
-
-
-
-
-
2
-
6
-
-
1
4
-
-
-
-
6
-
-
-
-
-
-
-
3
-
-
-
-
-
-
1
-
1
-
2
-
-
-
-
-
-
-
2
-
-
-
1
-
-
-
-
6
-
-
-
-
-
-
-
3
-
-
1
1
-
-
-
717241
Ouellet
Reaction of Mycobacterium tube ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Biochemistry
48
863-872
2009
-
-
1
-
-
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1
-
-
-
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-
-
161
-
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1
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1
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1
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1
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