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Literature summary extracted from

  • Frank, D.J.; Waddling, C.A.; La, M.; Ortiz de Montellano, P.R.
    Cytochrome P450 125A4, the third cholesterol C-26 hydroxylase from Mycobacterium smegmatis (2015), Biochemistry, 54, 6909-6916 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.29
-
 Mycolicibacterium smegmatis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.15.29 homology modeling of structure Mycolicibacterium smegmatis
1.14.15.29 to 2.26 A resolution Mycolicibacterium smegmatis

Protein Variants

EC Number Protein Variants Comment 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 Comment 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 UniProt Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.29 7alpha-hydroxycholest-4-en-3-one + 6 reduced ferredoxin [iron-sulfur] cluster + 6 H+ + 3 O2
-
 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
-
 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
-
 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
-
 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
-
 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
-
 Mycolicibacterium smegmatis ATCC 700084 (25S)-3beta-hydroxycholest-5-en-26-oate + 6 oxidized [2Fe-2S] ferredoxin + 4 H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.15.29 CYP125A3
-
 Mycolicibacterium smegmatis
1.14.15.29 CYP125A4
-
 Mycolicibacterium smegmatis

Expression

EC Number Organism Comment Expression
1.14.15.29 Mycolicibacterium smegmatis expression is not upregulated in response to cholesterol additional information