Information on EC 1.14.99.B1 - campesterol 22-hydroxylase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.99.B1
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
campesterol 22-hydroxylase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
campesterol + AH2 + O2 = 22alpha-hydroxycampesterol + A + H2O
show the reaction diagram
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SYSTEMATIC NAME
IUBMB Comments
campesterol,hydrogen donor:oxygen oxidoreductase (22-hydroxylating)
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CAS REGISTRY NUMBER
COMMENTARY hide
83380-86-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
cv. Momotarou
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
the expression of DWARF4, which encodes a C-22 hydroxylase, is crucial for brassinosteroid biosynthesis and for the feedback control of endogenous brassinosteroid levels. The amount of the DWF4 mRNA precursor either decreases or increases, similarly with its mature form, in response to an exogenously applied bioactive brassinosteroid, brassinolide, and a brassinosteroid biosynthesis inhibitor, brassinazole, respectively. Brassinosteroids act positively on root elongation under the control of auxin
physiological function
the expression of DWARF4, which encodes a C-22 hydroxylase, is crucial for brassinosteroid biosynthesis and for the feedback control of endogenous brassinosteroid levels. DWF4 transcription plays an important role in the brassinosteroid-auxin crosstalk associated with root elongation, in addition to its role in brassinosteroid homeostasis
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(24R)-ergost-4-en-3-one + NADPH + H+ + O2
(22S,24R)-22-hydroxy-ergost-4-en-3-one + NADP+ + H2O
show the reaction diagram
2,22-dideoxyecdysone + NADPH + H+ + O2
? + NADP+ + H2O
show the reaction diagram
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?
campestanol + NADPH + H+ + O2
6-deoxocathasterone + NADP+ + H2O
show the reaction diagram
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catalytic efficiency for campesterol is 325 times greater than for campestanol. 17% of the activity with cholesterol
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?
campesterol + NADPH + H+ + O2
(22S)-22-hydroxycampesterol + NADP+ + H2O
show the reaction diagram
best substrate
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-
?
campesterol + NADPH + H+ + O2
22alpha-hydroxycampesterol + NADP+ + H2O
show the reaction diagram
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catalytic efficiency for campesterol is 325 times greater than for campestanol. 76.9% of the activity with cholesterol
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?
cholestanol + NADPH + H+ + O2
22-hydroxy-cholestanol + NADP+ + H2O
show the reaction diagram
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29% of the activity with cholesterol
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?
cholesterol + NADPH + H+ + O2
22-hydroxy-cholesterol + NADP+ + H2O
show the reaction diagram
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?
sitosterol + NADPH + H+ + O2
22-hydroxy-sitosterol + NADP+ + H2O
show the reaction diagram
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poor substrate. 10.7% of the activity with cholesterol
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome P450
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NADH
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20% of the efficiency of NADPH plus a regenerating system
NADPH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
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heme iron
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2RS,4RS)-1-[4-propyl-2-(4-trifluoromethylphenyl)-1,3-dioxolan-2-ylmethyl]-1H-1,2,4-triazole
DWF4 is the target site of Brz220 in Arabidopsis. Arabidopsis thaliana treated with 2RS,4RS-1-[4-propyl-2-(4-trifluoromethylphenyl)-1,3-dioxane-2-ylmethyl]-1H-1,2,4-triazole shows a dwarf phenotype
2,22,25-trideoxyecdysone
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competitive
2-deoxyecdysone
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competitive
bitertanol
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i.e. 2,3,3-trimethyl-1-biphenyl4-yloxy-1-(1H-1,2,4-triazol-1-yl)butan-2-ol, Kd value is 0.0467 mM
brassinazole
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addition of brassinazole to the DWF4 protein induces a type II absorbance shift of the heme Soret band from 416 to 421 nm indicative of direct coordination of the triazole group of the brassinazole to the heme iron of DWF4. Kd value for brassinazole is 0.000001 M
Brz117
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i.e. 4-(3-methoxyphenyl)-2-phenyl-3-(1H-1,2,4-triazol-1-yl)butan-2-ol, Kd value is 0.0117 mM
difenoconazole
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Kd value is 0.0453 mM
ecdysone
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competitive
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0015
2,22-dideoxyecdysone
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pH 7.2, 37C
0.003
campestanol
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pH 7.25, 30C
0.00035
campesterol
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pH 7.25, 30C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00082
campestanol
Arabidopsis thaliana
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pH 7.25, 30C
0.03
campesterol
Arabidopsis thaliana
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pH 7.25, 30C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003
2,22,25-tridoexyecdysone
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pH 7.2, 37C
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0.006
2-deoxyecdysone
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pH 7.2, 37C
0.007
ecdysone
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pH 7.2, 37C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
transcript in mature fruit and in immature fruit; transcript in mature fruit, no transcript detected in immature fruit
Manually annotated by BRENDA team
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DWF4 expression is highest in apical shoots, and high in siliques. The expression of DWF4 in roots is comparable with that in shoots
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 55000, SDS-PAGE; x * 56000, SDS-PAGE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Arabidopsis plants ectopically overexpressing DWF4 (AOD4) are generated, using the cauliflower mosaic virus 35S promoter, and their phenotypes are characterized. The hypocotyl length of both light and dark-grown AOD4 seedlings is increased dramatically as compared to wild type. At maturity, inflorescence height increases >35% in AOD4 lines and >14% in tobacco DWF4 overexpressing lines (TOD4), relative to controls. The total number of branches and siliques increased more than twofold in AOD4 plants, leading to a 59% increase in the number of seeds produced
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expression in Escherichia coli
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expression in insect cell using a baculovirus expression system; expression in insect cell using a baculovirus expression system
gene DWF4, semi-quantitative reverse-transcriptase PCR expression analysis
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
downregulation of enzyme expression by brassinosteroid biosynthesis inhibitor brassinazole
upreguation by bioactive brassinosteroid, brassinolide
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
construction of DWF4::GUS plants, enzyme expression patterns in response to brassinazole and brassinolide in roots, overview
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
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it will be possible to control plant growth by engineering DWF4 transcription in plants