Information on EC 1.14.13.70 - sterol 14alpha-demethylase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.70
transferred to EC 1.14.14.154
RECOMMENDED NAME
GeneOntology No.
sterol 14alpha-demethylase
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Steroid biosynthesis
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Metabolic pathways
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Biosynthesis of secondary metabolites
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Biosynthesis of antibiotics
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
14alpha-methylzymosterol + NADPH + H+ + O2
?
show the reaction diagram
eburicol + NADPH + H+ + O2
?
show the reaction diagram
lanosterol + NADPH + H+ + O2
?
show the reaction diagram
obtusifoliol + NADPH + H+ + O2
?
show the reaction diagram
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol
i.e. oteseconazole, i.e. VT-1161. 98% inhibition, molar ratio of enzyme/inhibitor/lanosterol was 1:2:50
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(R)-N-(1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethyl)-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-benzamide
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(R)-N-(2-(1H-imidazol-1-yl)-1-phenylethyl)-4'-chlorobiphenyl-4-carboxamide
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1-(4-methyl-2-(4-(2-methylthiazol-4-yl)phenyl)thiazol-5-yl)ethanone
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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2,2-dimethyl-1,3-dichloropropane
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2-(methylamino)thiazol-4(5H)-one
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2-(naphthalen-1-ylamino)thiazol-4(5H)-one
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2-hydroxy-5-(2-(4-(2-methylthiazol-4-yl)phenyl)thiazol-4-yl)benzamide
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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2-methyl-4-(4-(4-(naphthalen-1-yl)thiazol-2-yl)phenyl)-thiazole
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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2-methyl-4-(4-(4-phenylthiazol-2-yl)phenyl)thiazole
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the compounds is predicted to be a non-competitive inhibitors of the enzyme by molecular docking study
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3-(alpha-imidazolylbenzyl)indole
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3-(alpha-triazolylbenzyl)indole
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4-(2-(4-(2-methylthiazol-4-yl)phenyl)thiazol-4-yl)-benzonitrile
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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4-(4-chlorophenyl)-2-(4-(2-methylthiazol-4-yl)phenyl)-thiazole
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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4-(4-methoxyphenyl)-2-(4-(2-methylthiazol-4-yl)-phenyl)thiazole
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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4-(chloromethyl)-2-(4-(2-methylthiazol-4-yl)phenyl)-thiazole
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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5-(2,4-dichlorobenzylidene)-2-(naphthalen-1-ylamino)thiazol-4(5H)-one
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5-(4-nitrobenzylidene)-2-(phenylamino)thiazol-4(5H)-one
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bifonazole
clotrimazole
difenoconazole
whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site
econazole
ethyl 2-(2-(4-(2-methylthiazol-4-yl)phenyl)thiazol-4-yl)acetate
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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ethyl 4-methyl-2-(4-(2-methylthiazol-4-yl)phenyl)-thiazole-5-carboxylate
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the compounds is predicted to be a non-competitive inhibitor of the enzyme by molecular docking study
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fluconazole
fluquinconazole
whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site
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itraconazole
ketaminazole
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ketoconazole
meglumine antimoniate
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miconazole
nitric oxide
in Huh7 human hepatoma cells, treatment with nitric oxide donors causes rapid post-translational down-regulation of the enzyme
oxo-prothioconazole
whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site
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posaconazole
Prochloraz
whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site
Propiconazole
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prothioconazole
whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site
prothioconazole-desthio
whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site
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tebuconazole
whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site
voriconazole
additional information
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the enzyme constitutes an important biological target for the most popular class of antifungals. Inhibition of lanosterol 14alpha-demethylase leads to accumulation of 14-a-methylsterols on the fungal surface and alteration of the permeability and rigidity of the plasma membrane, which results in arrest of fungal growth. Because this enzyme is found in all eukaryotes (including humans) and because the azoles interact also with other cytochrome P450 dependent enzymes (CYP3A4), a selective inhibition towards the fungal CYP51A1 is essential for an increased therapeutic index
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0559
eburicol
37C, pH 7.2
0.0063 - 0.0551
lanosterol
0.0421
obtusifoliol
37C, pH 7.2
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.09
eburicol
37C, PH 7.2
0.03 - 0.55
lanosterol
0.06
obtusifoliol
37C, PH 7.2
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.6
eburicol
37C, pH 7.2
0.54 - 87.3
lanosterol
1.4
obtusifoliol
37C, pH 7.2
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0044
3-(alpha-imidazolylbenzyl)indole
Leishmania mexicana
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pH and temperature not specified in the publication, axenic amastigotes
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0.00897
bifonazole
Leishmania infantum
A2TEF2
pH and temperature not specified in the publication
0.00281
clotrimazole
Leishmania infantum
A2TEF2
pH and temperature not specified in the publication
0.00536
econazole
Leishmania infantum
A2TEF2
pH and temperature not specified in the publication
0.206
fluconazole
Malassezia globosa
A8Q3I7
37C, pH 7.2
0.188
itraconazole
Malassezia globosa
A8Q3I7
37C, pH 7.2
0.321
ketaminazole
Malassezia globosa
A8Q3I7
37C, pH 7.2
-
0.176
ketoconazole
Malassezia globosa
A8Q3I7
37C, pH 7.2
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystals are grown using a hanging drop vapor diffusion technique. Enzyme complexes with posaconazole and VT-1161 are crystallized in the monoclinic C121 space group, and the structures are refined to 2.86 and 2.0 A. X-ray structures of Candida albicans CYP51 complexes with posaconazole and VT-1161, providing a molecular mechanism for the potencies of these drugs
X-ray crystal structures of hexahistidine-tagged Saccharomyces cerevisiae lanosterol 14alpha-demethylase in complex with its substrate lanosterol, the pseudosubstrate estriol and the triazole drugs itraconazole, posaconazole, fluconazole and voriconazole
for crystallization purposes, an N-terminal truncated construct is used where the membrane anchor sequence is replaced with the 5-amino acid sequence fragment MAKKT-. Crystals are grown at 23C using the hanging-drop vapor diffusion method
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression in Escherichia coli strain HMS174(DE3)
expression of mutant and wild-type enzymes in Pichia pastoris
recombinant hexahistidine-tagged enzyme is overexpresssed in a yeast membrane protein expression system
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N136Y
N136Y transformants show a reduced in vitro susceptibility to fluconazole compared to wild-type controls. The amino acid substitution N136Y in Candida albicans sterol 14alpha-demethylase is involved in fluconazole resistance
Y140F
3.3fold reduction in susceptibility to S-prothioconazole
Y140H
4.3fold reduction in susceptibility to S-prothioconazole
Y140F
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3.3fold reduction in susceptibility to S-prothioconazole
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Y140H
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4.3fold reduction in susceptibility to S-prothioconazole
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
pharmacology