BRENDA - Enzyme Database

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)
EC Number
Commentary
Organism
1.14.15.29
;
Mycolicibacterium smegmatis
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.14.15.29
homology modeling of structure; to 2.26 A resolution
Mycolicibacterium smegmatis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.14.15.29
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
1.14.15.29
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]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.15.29
0.0041
-
7alpha-hydroxycholest-4-en-3-one
wild-type, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
1.14.15.29
0.0051
-
7alpha-hydroxycholest-4-en-3-one
mutant Y87W, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
1.14.15.29
0.0241
-
cholest-4-en-3-one
mutant W87Y, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.15.29
Mycolicibacterium smegmatis
A0QY38
-
-
1.14.15.29
Mycolicibacterium smegmatis
A0R4Y3
-
-
1.14.15.29
Mycolicibacterium smegmatis ATCC 700084
A0QY38
-
-
1.14.15.29
Mycolicibacterium smegmatis ATCC 700084
A0R4Y3
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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)
EC Number
Commentary
Organism
1.14.15.29
-
Mycolicibacterium smegmatis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.14.15.29
homology modeling of structure
Mycolicibacterium smegmatis
1.14.15.29
to 2.26 A resolution
Mycolicibacterium smegmatis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.14.15.29
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
1.14.15.29
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)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.15.29
0.0041
-
7alpha-hydroxycholest-4-en-3-one
wild-type, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
1.14.15.29
0.0051
-
7alpha-hydroxycholest-4-en-3-one
mutant Y87W, pH 7.4, temperature not specified in the publication
Mycolicibacterium smegmatis
1.14.15.29
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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
-
-
-
?
1.14.15.29
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
EC Number
Organism
Commentary
Expression
1.14.15.29
Mycolicibacterium smegmatis
expression is not upregulated in response to cholesterol
additional information
Expression (protein specific)
EC Number
Organism
Commentary
Expression
1.14.15.29
Mycolicibacterium smegmatis
expression is not upregulated in response to cholesterol
additional information