BRENDA - Enzyme Database

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)
EC Number
Commentary
Organism
1.14.15.28
-
Mycobacterium tuberculosis
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.14.15.28
sitting drop method, the CYP142 crystal structure is solved to 1.6 A
Mycobacterium tuberculosis
General Stability
EC Number
General Stability
Organism
1.14.15.28
completely to the P450 state on binding of cholest-4-en-3-one at pH 8.0
Mycobacterium tuberculosis
1.14.15.28
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)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.15.28
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
-
-
?
1.14.15.28
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.15.28
Mycobacterium tuberculosis
P9WPL5
-
-
1.14.15.28
Mycobacterium tuberculosis ATCC 25618
P9WPL5
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.15.28
-
Mycobacterium tuberculosis
Reaction
EC Number
Reaction
Commentary
Organism
1.14.15.28
(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
1.14.15.28
(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
1.14.15.28
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
1.14.15.28
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
1.14.15.28
6
-
the substrate free enzyme is unstable and aggregates
Mycobacterium tuberculosis
1.14.15.28
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
1.14.15.28
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)
EC Number
Commentary
Organism
1.14.15.28
-
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.14.15.28
sitting drop method, the CYP142 crystal structure is solved to 1.6 A
Mycobacterium tuberculosis
General Stability (protein specific)
EC Number
General Stability
Organism
1.14.15.28
completely to the P450 state on binding of cholest-4-en-3-one at pH 8.0
Mycobacterium tuberculosis
1.14.15.28
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)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.15.28
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
-
-
?
1.14.15.28
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)
EC Number
Commentary
Organism
1.14.15.28
-
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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
-
-
-
?
1.14.15.28
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)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
1.14.15.28
6
-
the substrate free enzyme is unstable and aggregates
Mycobacterium tuberculosis
1.14.15.28
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
1.14.15.28
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
EC Number
General Information
Commentary
Organism
1.14.15.28
metabolism
the enzyme is involved in host response modulation and cholesterol metabolism
Mycobacterium tuberculosis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.14.15.28
metabolism
the enzyme is involved in host response modulation and cholesterol metabolism
Mycobacterium tuberculosis