BRENDA - Enzyme Database
show all sequences of 1.14.15.29

Cytochrome P450 125A4, the third cholesterol C-26 hydroxylase from Mycobacterium smegmatis

Frank, D.J.; Waddling, C.A.; La, M.; Ortiz de Montellano, P.R.; Biochemistry 54, 6909-6916 (2015)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
;
Mycolicibacterium smegmatis
Crystallization (Commentary)
Crystallization
Organism
homology modeling of structure; to 2.26 A resolution
Mycolicibacterium smegmatis
Engineering
Amino acid exchange
Commentary
Organism
W83Y
mutation to the corresponding residue of isoform CYP125A4. Mutation reduces the size of the binding pocket, reducing the ability to bind 7alpha-hydroxylated substrates
Mycolicibacterium smegmatis
Y83W
mutation to the corresponding residue of isoform CYP125A3. Mutation increases the size of the binding pocket, increasing the ability to bind 7alpha-hydroxylated substrates. Enzyme shows activity toward 4-cholesten-3-one, albeit with sigmoidal kinetics
Mycolicibacterium smegmatis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0041
-
7alpha-hydroxycholest-4-en-3-one
wild-type, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
0.0051
-
7alpha-hydroxycholest-4-en-3-one
mutant Y87W, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
0.0241
-
cholest-4-en-3-one
mutant W87Y, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mycolicibacterium smegmatis
A0QY38
-
-
Mycolicibacterium smegmatis
A0R4Y3
-
-
Mycolicibacterium smegmatis ATCC 700084
A0QY38
-
-
Mycolicibacterium smegmatis ATCC 700084
A0R4Y3
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7alpha-hydroxycholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis
(25S)-7alphahydroxy-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
7alpha-hydroxycholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis ATCC 700084
(25S)-7alphahydroxy-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis
(25S)-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis ATCC 700084
(25S)-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
cholest-5-en-3beta-ol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
744344
Mycolicibacterium smegmatis
(25S)-3beta-hydroxycholest-5-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-5-en-3beta-ol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
744344
Mycolicibacterium smegmatis ATCC 700084
(25S)-3beta-hydroxycholest-5-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Mycolicibacterium smegmatis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
homology modeling of structure
Mycolicibacterium smegmatis
to 2.26 A resolution
Mycolicibacterium smegmatis
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
W83Y
mutation to the corresponding residue of isoform CYP125A4. Mutation reduces the size of the binding pocket, reducing the ability to bind 7alpha-hydroxylated substrates
Mycolicibacterium smegmatis
Y83W
mutation to the corresponding residue of isoform CYP125A3. Mutation increases the size of the binding pocket, increasing the ability to bind 7alpha-hydroxylated substrates. Enzyme shows activity toward 4-cholesten-3-one, albeit with sigmoidal kinetics
Mycolicibacterium smegmatis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0041
-
7alpha-hydroxycholest-4-en-3-one
wild-type, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
0.0051
-
7alpha-hydroxycholest-4-en-3-one
mutant Y87W, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
0.0241
-
cholest-4-en-3-one
mutant W87Y, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7alpha-hydroxycholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis
(25S)-7alphahydroxy-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
7alpha-hydroxycholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis ATCC 700084
(25S)-7alphahydroxy-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis
(25S)-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
cholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
744344
Mycolicibacterium smegmatis ATCC 700084
(25S)-3-oxocholest-4-en-26-oate + 6 oxidized ferredoxin [iron-sulfur] cluster + 4 H2O
-
-
-
?
cholest-5-en-3beta-ol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
744344
Mycolicibacterium smegmatis
(25S)-3beta-hydroxycholest-5-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
cholest-5-en-3beta-ol + 6 reduced [2Fe-2S] ferredoxin + 3 O2
-
744344
Mycolicibacterium smegmatis ATCC 700084
(25S)-3beta-hydroxycholest-5-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
-
-
?
Expression
Organism
Commentary
Expression
Mycolicibacterium smegmatis
expression is not upregulated in response to cholesterol
additional information
Expression (protein specific)
Organism
Commentary
Expression
Mycolicibacterium smegmatis
expression is not upregulated in response to cholesterol
additional information
Other publictions for EC 1.14.15.29
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)
744344
Frank
Cytochrome P450 125A4, the th ...
Mycolicibacterium smegmatis, Mycolicibacterium smegmatis ATCC 700084
Biochemistry
54
6909-6916
2015
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-
1
1
2
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3
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6
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12
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2
-
2
2
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3
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12
-
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-
-
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-
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-
1
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1
-
-
744481
Johnston
Substrate analog studies of t ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Bioorg. Med. Chem.
20
4064-4081
2012
-
-
-
-
-
-
-
-
-
-
-
4
-
4
-
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-
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8
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3
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4
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8
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3
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-
-
-
-
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717344
Ouellet
Reverse type I inhibitor of My ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Bioorg. Med. Chem. Lett.
21
332-337
2011
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-
-
1
1
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1
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6
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2
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1
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1
1
1
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1
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2
-
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-
717811
Johnston
Functional redundancy of stero ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis CDC 1551, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
285
36352-36360
2010
-
-
1
-
-
-
-
2
-
-
-
-
-
165
-
-
1
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6
-
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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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6
-
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2
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-
-
-
-
-
-
718129
Ouellet
Mycobacterium tuberculosis CYP ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis CDC 1551
Mol. Microbiol.
77
730-742
2010
-
-
1
1
-
-
-
-
-
-
-
2
-
4
-
-
1
-
-
-
-
-
2
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-
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-
1
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1
1
1
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2
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1
-
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-
-
2
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
717241
Ouellet
Reaction of Mycobacterium tube ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Biochemistry
48
863-872
2009
-
-
1
-
-
-
1
-
-
-
-
-
-
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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-
-
-
-
717782
McLean
The structure of Mycobacterium ...
Mycobacterium tuberculosis
J. Biol. Chem.
284
35524-35533
2009
-
-
1
1
-
-
-
-
-
-
-
-
-
3
-
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1
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1
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2
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1
2
1
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1
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1
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717783
Capyk
Mycobacterial cytochrome P450 ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv, Mycobacterium tuberculosis variant bovis, Mycobacterium tuberculosis variant bovis bacillus Calmette-Guerin
J. Biol. Chem.
284
35534-35542
2009
-
-
2
-
-
-
-
1
-
-
2
-
-
168
-
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2
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8
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2
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2
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2
2
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1
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2
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8
-
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2
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-
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1
2
2
1
1
1
718126
Rosloniec
Cytochrome P450 125 (CYP125) c ...
Rhodococcus jostii
Mol. Microbiol.
74
1031-1043
2009
-
-
1
-
-
-
-
-
-
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1
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5
-
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1
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1
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2
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1
2
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1
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1
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1
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2
2
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