Information on EC 1.13.11.27 - 4-hydroxyphenylpyruvate dioxygenase

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The expected taxonomic range for this enzyme is: Eukaryota, Archaea, Bacteria

EC NUMBER
COMMENTARY hide
1.13.11.27
-
RECOMMENDED NAME
GeneOntology No.
4-hydroxyphenylpyruvate dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decarboxylation
-
-
-
-
hydroxylation
-
-
-
-
redox reaction
-
-
-
-
side-chain migration
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-phenylalanine biosynthesis III (cytosolic, plants)
-
-
L-phenylalanine degradation IV (mammalian, via side chain)
-
-
L-tyrosine degradation I
-
-
Metabolic pathways
-
-
Phenylalanine metabolism
-
-
plastoquinol-9 biosynthesis I
-
-
tyrosine metabolism
-
-
Tyrosine metabolism
-
-
Ubiquinone and other terpenoid-quinone biosynthesis
-
-
vitamin E biosynthesis (tocopherols)
-
-
SYSTEMATIC NAME
IUBMB Comments
4-hydroxyphenylpyruvate:oxygen oxidoreductase (hydroxylating, decarboxylating)
The Pseudomonas enzyme contains one Fe3+ per mole of enzyme; the enzymes from other sources may contain essential iron or copper.
CAS REGISTRY NUMBER
COMMENTARY hide
9029-72-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Frog
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Pseudomonas sp. P.J. 874
strain P.J. 874
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain MR-1
UniProt
Manually annotated by BRENDA team
Vitis vinifera x Vitis riparia
-
-
-
Manually annotated by BRENDA team
Vitis vinifera x Vitis vinifera
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
deficiency in active 4-HPPD in humans results in type III tyrosinemia, a rare autosomal recessive disorder
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2-fluoro-4-hydroxyphenyl)pyruvate + O2
(3-fluoro-2,5-dihydroxyphenyl)pyruvate + CO2
show the reaction diagram
(4-hydroxyphenyl)-pyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
2-thienylpyruvate + O2
? + CO2
show the reaction diagram
-
-
-
-
?
3,4-dihydroxyphenylpyruvate + O2
? + CO2
show the reaction diagram
3-thienylpyruvate + O2
3-carboxymethyl-3-thiolene-2-one + CO2
show the reaction diagram
-
-
-
?
4-fluorophenylpyruvate + O2
4-fluoro-2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
2,5-dihydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
4-hydroxyphenylpyruvate + O2
homogentisate + CO2 + 4-hydroxyphenylacetate
show the reaction diagram
4-hydroxytetrafluorophenylpyruvate + O2
? + CO2
show the reaction diagram
-
-
-
-
?
4-methylphenylpyruvate + O2
? + CO2
show the reaction diagram
-
poor substrate
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
[(4-hydroxyphenyl)thio]pyruvate + O2
[(4-hydroxyphenyl)sulfinyl]acetate + CO2
show the reaction diagram
additional information
?
-
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
4-hydroxyphenylpyruvate + O2
2,5-dihydroxyphenylacetate + CO2
show the reaction diagram
P80064
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
4-hydroxyphenylpyruvate + O2
homogentisate + CO2 + 4-hydroxyphenylacetate
show the reaction diagram
additional information
?
-
-
HpdR positively regulates hpdA expression through direct binding to the HpdA promoter within a region containing two conserved direct repeat sequences. HpdR-dependent hpdA transcription occurs in the presence of 4-hydroxyphenylpyruvate, tyrosine, and phenylalanine, as well as during starvation
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
copper
Zinc
-
0.68 mol of zinc per mol of enzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2,6-difluoro-4-hydroxyphenyl)pyruvate
-
competitive, most potent
1,10-phenanthroline
1,2-dihydroxybenzene
-
0.019 mM, 50% inhibition, uncompetitive
1,2-dihydroxybenzene-3,5-disulfonic acid
-
inhibition is prevented by ascorbate but not reversed by enol-4-hydroxyphenylpyruvate
1,4-benzoquinone
-
0.25 mM, 50% inhibition, competitive
1,4-dihydroxybenzene
-
4.7 mM, 50% inhibition
1-(1,1-dimethylethyl)-5-hydroxy-4-(3-(4-(methoxy)phenyl)-2-methyl-4-(methylsulfonyl)benzoyl)pyrazole
1-(2'-theonyl)-2,3-trifluoroacetone
-
0.0049 mM, 50% inhibition
-
1-(2'-thionyl)-3,3,3-trifluoroacetone
-
-
2,2'-bipyridyl
2,4-Dihydroxybenzoate
-
4.2 mM, 50% inhibition, noncompetitive
2,4-Dihydroxyphenylpyruvate
-
competitive
2,5-Dihydroxybenzoate
-
2.6 mM, 50% inhibition, noncompetitive
2,5-dihydroxyphenylacetate
-
50% inhibition above 10 mM, noncompetitive
2,5-dihydroxyphenylacetic acid lactone
-
50% inhibition above 10 mM, competitive
2-(2-chloro-4-methane sulfonylbenzoyl)-1,3-cyclohexanedione
competitive inhibitor
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione
2-(2-nitro-4-(trifluoromethyl)benzoyl)-1,3-cyclohexanedione
-
(NTBC), binding constant of 937 microM
2-(2-nitro-4-chlorobenzoyl)-5-(2-methoxyethyl)cyclohexane-1,3-dione
-
-
2-Hydroxybenzaldehyde
-
competitive
2-hydroxyphenylacetate
2-Hydroxyphenylpyruvate
-
competitive
2-mercaptoethanol
10 mM, 65% inhibition
2-thienylpyruvate
-
mechanism-based inhibitor
2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
2-[2-nitro-4-(trifluoromethyl)benzoyl]-4,4,6,6-tetramethylcyclohexane-1,3,5-trione
-
50 nM, 50% inhibition, tight-binding reversible inhibitor, rapid inactivation by the formation of an enzyme-inhibitor complex that dissociates extremely slowly with recovery of enzyme activity
3,3,5,5-tetramethyl-2,4,6-trioxocyclohexyl 3-methylbutanoate
-
-
3,4-dihydroxybenzoate
-
0.24 mM, 50% inhibition, noncompetitive
3,4-Dihydroxyphenylacetate
-
0.75 mM, 50% inhibition, noncompetitive
3,5-diiodo-4-hydroxyphenylpyruvate
-
0.25 mM, 50% inhibition, competitive
3-(2-cyclohexylethyl)-5,5,7,7-tetramethylbenzo[d]isoxazole-4,6(5H,7H)-dione
-
-
3-(2-hydroxyphenyl)propionate
-
50% inhibition above 10 mM, competitive
3-cyclohexyl-1-(2,4,6-trihydroxy-3-(3-methylbut-2-enyl)phenyl)propan-1-one
-
-
3-hydroxy-4-phenyl-2-furanone
-
0.0005 mM, 50% inhibition, may serve as a lead compound for further design of more potent inhibitors
3-hydroxyphenylpyruvate
-
competitive
3-methoxy-4-hydroxyphenylacetate
-
0.25 mM, 50% inhibition, competitive
3-methoxy-4-hydroxyphenylpyruvate
-
competitive
3-thienylpyruvate
-
mechanism-based inhibitor
4-(2,4-dichlorobenzyl)-1,3-dimethyl-5-hydroxypyrazole
-
4-(3-cyclohexylpropanoyl)-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
4-(3-cyclohexylpropanoyl)-5-hydroxy-2,2,6-trimethyl-6-(3-methylbut-2-enyl)cyclohex-4-ene-1,3-dione
-
-
4-(3-cyclohexylpropanoyl)-5-methoxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
4-(cyclohexanecarbonyl)-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
4-acetyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
4-aminophenylpyruvate
-
competitive
4-decanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
4-dodecanoyl-5-hydroxy-2,2,6,6-tetrakis(3-methylbut-2-enyl)cyclohex-4-ene-1,3-dione
-
-
4-dodecanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
4-hexanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
4-hydroxybenzaldehyde
-
50% inhibition above 10 mM, noncompetitive
4-hydroxyphenylpyruvate
4-hydroxytetrafluorophenylpyruvate
-
mechanism-based inhibitor
5-hydroxy-2,2,6,6-tetramethyl-4-(2-methylbutanoyl)cyclohex-4-ene-1,3-dione
-
-
5-hydroxy-2,2,6,6-tetramethyl-4-(2-phenylacetyl)cyclohex-4-ene-1,3-dione
-
-
5-hydroxy-2,2,6,6-tetramethyl-4-(3-methylbutanoyl)cyclohex-4-ene-1,3-dione
-
-
5-hydroxy-2,2,6,6-tetramethyl-4-(3-phenylpropanoyl)cyclohex-4-ene-1,3-dione
-
-
5-hydroxy-2,2,6,6-tetramethyl-4-palmitoylcyclohex-4-ene-1,3-dione
-
-
5-hydroxy-4-isobutyryl-2,2,6,6-tetrakis(3-methylbut-2-enyl)cyclohex-4-ene-1,3-dione
-
-
5-hydroxy-4-isobutyryl-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
8-Hydroxyquinoline-5-sulfonic acid
-
560 nM, 50% inhibition
Acetopyruvate
-
0.33 mM, 50% inhibition, competitive
Al3+
10 mM, 7% inhibition
Bathocuproine
-
3 mM, 50% inhibition
bathophenanthroline
benzobicyclon hydrolysate
-
Ca2+
10 mM, 16% inhibition
citrate
-
-
Co2+
10 mM, 32% inhibition
Cu2+
10 mM, complete inhibition
Cupferron
diethyl dicarbonate
-
5 mM, 50% inhibition
diethyldithiocarbamate
diketonitrile of isoxaflutole
-
half-site reactivity, nearly irreversible
Dithizone
DL-3,4-dihydroxyphenylalanine
-
0.32 mM, 50% inhibition, noncompetitive
DL-4-hydroxyphenyllactate
-
50% inhibition above 10 mM, competitive
DL-Epinephrine
-
0.17 mM, 50% inhibition, noncompetitive
DTT
10 mM, 61% inhibition
ethanol
-
5% v/v, 50% inhibition, complete loss of activity at 20% v/v
Fe2+-chelators
-
-
-
homogentisate
K+
10 mM, 6% inhibition
L-Phe
-
10 mM, 50% inhibition, competitive
LnCl3
-
1 mM, 50% inhibition
-
mesotrione
-
binding constant of 602 microM
Mg2+
10 mM, 18% inhibition
Mn2+
10 mM, 97% inhibition
Na+
10 mM, 14% inhibition
Ni2+
10 mM, 28% inhibition
o-hydroxyphenylpyruvate
-
competitive inhibitor of 4-hydroxyphenylpyruvate
oxaloacetate
-
10 mM, 50% inhibition, noncompetitive
pentafluorophenylpyruvate
-
competitive
phenylglyoxalate
-
50% inhibition above 10 mM, competitive
phenylpyruvate
pyridoxal 5'-phosphate
-
2 mM, 50% inhibition
pyruvate
-
50% inhibition above 10 mM, competitive
SDS
2% (w/v), complete inhibition
sulcotrione
for protein expressed in Escherichia coli DH5alpha (inhibition assayed without preincubation): 0 micoM sulcotrione = 100% activity, 0.5 microM sulcotrione = 58% activity left, 1.0 microM sulcotrione = 21.3% activity left, 1.5 microM sulcotrione = 2.8% activity left. Preincubation of the enzyme expressed in Escherichia coli SG13009 with sulcotrione for 15 min, significantly enhances the inhibition from 40% (1 microM sulcotrione/no preincubation) to 10% (1 microM sulcotrione/15 min preincubation).
thiophenol oxalate
-
-
thiophenyl oxalate
-
competitive
Tiron
-
0.4 mM, 50% inhibition
topramezone
-
-
trans-4-hydroxycinnamate
-
50% inhibition above 10 mM, competitive
triketone-rich fraction
-
-
Urea
1 M, 3% inhibition. 3 M, 42% inhibition. 5 M, 56% inhibition
YCl3
-
1 mM, 50% inhibition
Zn2+
10 mM, 65% inhibition
[1-tert-butyl-3-(2,4-dichlorophenyl)-5-hydroxy-1H-pyrazol-4-yl][2-chloro-4-(methylsulfonyl)phenyl]methanone
-
50% inhibition at 12 nM
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-butanol
-
5% v/v, 2.9fold activation
1-propanol
-
10% v/v, 2.1fold activation
2,6-dichlorophenol indophenol
acetone
-
10% v/v, 5.7fold activation
acetonitrile
-
10% v/v, 3.6fold activation
ascorbate
catalase
-
from bovine liver, required for activity
-
Cyclohexanol
-
5% v/v, 4.4fold activation
dioxane
-
10% v/v, 2.8fold activation
ethanol
-
10% v/v, 2fold activation
ether
-
10% v/v, 2.6fold activation
glutathione
-
reducing cofactor required, either ascorbate, or a combination of 2,6-dichlorophenol indophenol and glutathione in presence of catalase
methanol
-
10% v/v, 1.6fold activation
methyl-cellosolve
-
10% v/v, 2.2fold activation
tetrahydrofuran
-
10% v/v, 6.7fold activation
Trypsin
Frog
-
liver enzyme appears to be present in an inactive form which can be activated by treatment with trypsin or by autolysis
-
additional information
regarding expression and acitvity, rich or complex media (surplus of yeast extract, addition of glycerol and buffering of pH) are superior to media with less complex composition. Supplementation of media with FeSO4 or tyrosine does not substantially upgrade expression and activity levels.
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.027
(4-hydroxyphenyl)-pyruvate
-
pH 7.0, 25C
0.5
2-thienylpyruvate
-
-
0.05
3,4-Dihydroxyphenylpyruvate
-
-
0.25
3-thienylpyruvate
-
-
0.0043 - 54
4-hydroxyphenylpyruvate
0.05
4-hydroxytetrafluorophenylpyruvate
0.069 - 0.1
O2
0.115
oxygen
-
activity is measured polarographically, assays include 0.5 microM of enzyme, 500 microM betaME, Fe2+ stoichiometric with enzyme, and 400 microM 4-hydroxyophenylpyruvate in 20 mM HEPES at pH 7.0 at 4C
0.06 - 0.52
phenylpyruvate
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.8
(4-hydroxyphenyl)-pyruvate
Streptomyces avermitilis
-
-
0.03 - 9.9
4-hydroxyphenylpyruvate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.27 - 254
4-hydroxyphenylpyruvate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0013
(2,6-difluoro-4-hydroxyphenyl)pyruvate
-
-
0.0000098
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione
-
-
0.4 - 1.9
4-hydroxyphenylpyruvate
0.014
pentafluorophenylpyruvate
-
-
0.15
thiophenol oxalate
-
-
additional information
additional information
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00675
4-(2,4-dichlorobenzyl)-1,3-dimethyl-5-hydroxypyrazole
Coptis japonica var. dissecta
A7BG64
recombinant enzyme
0.00017
4-(3-cyclohexylpropanoyl)-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.00227
4-(3-cyclohexylpropanoyl)-5-hydroxy-2,2,6-trimethyl-6-(3-methylbut-2-enyl)cyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.00041
4-(3-cyclohexylpropanoyl)-5-methoxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.000019
4-decanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.00025
4-dodecanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.00096
4-hexanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.0143
5-hydroxy-2,2,6,6-tetramethyl-4-(2-methylbutanoyl)cyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.0189
5-hydroxy-2,2,6,6-tetramethyl-4-(2-phenylacetyl)cyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.0118
5-hydroxy-2,2,6,6-tetramethyl-4-(3-methylbutanoyl)cyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.00075
5-hydroxy-2,2,6,6-tetramethyl-4-(3-phenylpropanoyl)cyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
grandiflorone, most active naturally occurring beta-triketone
0.00031
5-hydroxy-2,2,6,6-tetramethyl-4-palmitoylcyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.0445
5-hydroxy-4-isobutyryl-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
Arabidopsis thaliana
-
-
0.00234
benzobicyclon hydrolysate
Coptis japonica var. dissecta
A7BG64
recombinant enzyme
0.000015 - 0.000023
topramezone
additional information
3,3,5,5-tetramethyl-2,4,6-trioxocyclohexyl 3-methylbutanoate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.004
protein expressed in Escherichia coli DH5alpha
0.04
purified recombinant mutant R378K, pH 6.5, 37C, homogentisate production
0.06
purified recombinant mutant E254D, pH 6.5, 37C, O2 consumption
0.07
purified recombinant mutant R378K, pH 6.5, 37C, O2 consumption
0.11 - 0.2
protein expressed in Escherichia coli SG13009
0.17 - 0.26
protein expressed in Escherichia coli M15(pRep4)
0.2
purified recombinant mutant E254D, pH 6.5, 37C, homogentisate production
0.51 - 1.1
-
enzyme forms 1, 2, and 3
0.53
-
-
0.6 - 0.9
-
-
2.6
purified recombinant wild-type enzyme, pH 6.5, 37C, O2 consumption
2.8
purified recombinant wild-type enzyme, pH 6.5, 37C, homogentisate production
10.6
-
pH 7.0, 25C
39.6
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
two pH optima are observed at 50C: pH 4.0 (close to native conditions, narrow maximum) and pH 7.0 (broad maximum)
4.5
-
and a second optimum at pH 7.8
4.6
-
and a second optimum at pH 7.4, enzyme form 3
5.1
-
and a second optimum at pH 7.4, enzyme form 1 and 2
6 - 7.5
-
-
7.3
-
-
7.4
-
and a second optimum at pH 5.1 for enzyme form 1 and 2, and at pH 4.6 for enzyme form 3
7.8
-
and a second optimum at pH 4.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7
-
reaction rate is constant between pH 5 and pH 7
5.5 - 7.7
-
pH 5.5: about 50% of maximal activity, pH 7.7: about 55% of maximal activity
6 - 9.5
-
pH 6.0: about 55% of maximal activity, pH 9.5: about 50% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
assay at
50
maximal activity when assayed at pH 4.0 or 7.0
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 70
30C: about 65% of maximal activity, 70C: about 30% of maximal activity, assayed at pH 7.0
40 - 60
40C: about 55% of maximal activity, 60C: about 40% of maximal activity, assayed at pH 4.0
additional information
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
abundantly expressed in neurons of
Manually annotated by BRENDA team
abundantly expressed in neurons of
Manually annotated by BRENDA team
-
etiolated
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
-
in transgenic Glycine max (cv. Jack), SDS-PAGE; in transgenic Nicotiana tabacum (cv. PBD6), Western blot
50000 - 54000
-
gel filtration
52540
predicted molecular mass
63000 - 67000
-
enzyme forms 1, 2, and 3, gel filtration
66000
-
gel filtration
85000
-
gel filtration
87000
-
equilibrium sedimentation
97000
-
equilibrium sedimentation
100000
-
non-denaturing PAGE
150000
-
high-speed equilibrium sedimentation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
-
recombinant enzyme can be phosphorylated in human AMA cells, phosphorylation is not indispensable for activity
proteolytic modification
undergoes both N-and C-terminal processing, C-terminal processing enhances enzymic activity
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
substrate-free form
-
with and without inhibitors [1-tert-butyl-3-(2,4-dichlorophenyl)-5-hydroxy-1H-pyrazol-4-yl][2-chloro-4-(methylsulfonyl)phenyl]methanone or 1-(1,1-dimethylethyl)-5-hydroxy-4-(3-(4-(methoxy)phenyl)-2-methyl-4-(methylsulfonyl)benzoyl)pyrazole
-
crystals are grown by sitting-drop vapor diffusion method at 20C
crystal structure at 2.4 A resolution
-
hanging-drop method in 18-25% PEG 4000, 0.2 M ammonium acetate, 0.1 M trisodium citrate, pH 5.6, at room temperature
-
with and without inhibitors 1-(1,1-dimethylethyl)-5-hydroxy-4-(3-(4-(methoxy)phenyl)-2-methyl-4-(methylsulfonyl)benzoyl)pyrazole or [1-tert-butyl-3-(2,4-dichlorophenyl)-5-hydroxy-1H-pyrazol-4-yl][2-chloro-4-(methylsulfonyl)phenyl]methanone
Fe(II)-form in complex with inhibitor 2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
-
substrate-free form
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.5 - 10
-
37C, 30 min, in presence of Fe2+, stable
395381
5.5 - 8
-
37C, 15 min, stable
395375
7.5
-
37C, 7.5 min, stable
395376
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
-20
enzyme activity is stable for a few weeks.
40
pH 4.0, half-life: 1.5 h
77
strong conformational changes are observed at 77C, indicating melting of the protein
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
long-term storage of concentrated solutions in presence of air leads to polymerization
-
regeneration of time-dependent inactivated 4-hydroxyphenylpyruvate dioxygenase by incubation with ascorbic acid and/or Fe2+ salts is not successful
stable to freezing and thawing
-
the enzyme is stable for several hours at room temperature, indefinitely at 4C
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
routinely stored in presence of O2
-
395377
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, concentrated enzyme solution, 1.0 mg/ml, stable for 6 months or more
-
-20C, concentrated solution, 1 mg/ml, pH 6.0, stable for 6 months or more
-
-20C, on long-term storage the enzyme forms polymers, reversal by thiols
-
-60C, 0.2 M NaCl solution buffered with 10 mM potassium phosphate, pH 6.7, stable for several months
-
-60C, pH 7.5, 3 months, 20% loss of activity when enzyme concentration is 0.5 g/L in Tris/HCl buffer, no loss of activity after 4 years when enzyme concentration is 5 g/l
-
-60C, stable for several months
4C, 10 mM sodium acetate buffer, pH 6.0, little loss of activity after 2 months
-
4C, dilute solutions, 0.1 mg/ml, 100 mM sodium acetate buffer, PH 6.0, stable for up to 2 months
-
storage at room temperature results in residual enzyme activities of ca. 60-80% after 16 h or 40-50% after 30 h.
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at 5C or below
-
immobilized metal affinity chromatography
-
Q Sepharose anion exchange column, Sephacryl S-200 gel filtration
-
recombinant enzyme 15fold from Escherichia coli by anion-exchange and hydrophobic interaction chromatography, and gel filtration
recombinant enzyme expressed in Escherichia coli
recombinant wild-type and mutant enzymes by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
-
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
barley 4-hydroxyphenylpyruvate dioxygenase gene introduced into the plastome of Nicotiana tabacum (tobacco)
DNA and amino acid sequence determination and analysis, human enzyme expression in Escherichia coli
expressed in Escherichia coli BL21 DE3
-
expressed in Eschrichia coli stains DH5alpha, M15(pRep4) or SG13009
expressed in Glycine max (cv. Jack); expressed in Nicotiana tabacum (cv. PBD6)
-
expressed in Nicotiana tabacum
-
expression in Escherichia coli
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
overexpressed in Escherichia coli
-
overexpression of Arabidopsis thaliana p-hydroxyphenylpyruvate dioxygenase and Arabidopsis thaliana homogentisate phytyltransferase in Solanum tuberosum. Over-expression of At-HPPD results in a maximum 266% increase in alpha-tocopherol, and over-expression of At-HPT yields a 106% increase. Tubers from transgenic plants accumulate approximately 10fold and 100fold less a-tocopherol than leaves or seeds, respectively
-
overproduction of the recombinant enzyme in Escherichia coli JM105, overexpression of the complete coding sequence in transgenic Nicotiana tabacum using Agrobacterium tumefaciens transformation
-
recombinant expression of wild-type and mutant enzymes
-
vaccinia virus-based expression in human AMA cells and expression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
for Ag+ treatment, Smhppd transcription level gradually decreases along with the prolongation of treatment duration (P less than 0.05), with a minimum of about threefold lower than that of control
methyl jasmonate and salicylic acid can induce Smhppd expression (P less than 0.05), and the highest level is both observed on 6 days after treatment, with more than twofold higher than that of control; Smhppd expression is gradually up-regulated (P less than 0.05), with a maximum of more than two times on 12 days after yeast extract treatment
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N261D
-
site-directed mutagenesis, during the reaction mechanism, the last 1,2 rearrangement is blocked in S246A HPPD mutant, so that an arene oxide-derived intermediate is released as an alternative product, the mutant shows increased Km and reduced kcat for 4-hydroxyphenylpyruvate compared to the wild-type enzyme
Q272E
-
site-directed mutagenesis, the mutant shows increased Km and reduced kcat for 4-hydroxyphenylpyruvate compared to the wild-type enzyme
Q286E
-
site-directed mutagenesis, the mutant shows increased Km and reduced kcat for 4-hydroxyphenylpyruvate compared to the wild-type enzyme
Q358E
-
site-directed mutagenesis, the mutant shows increased Km and reduced kcat for 4-hydroxyphenylpyruvate compared to the wild-type enzyme
S246A
-
site-directed mutagenesis, during the reaction mechanism, the last 1,2 rearrangement is blocked in S246A HPPD mutant, so that an arene oxide-derived intermediate is released as an alternative product, the mutant shows increased Km and reduced kcat for 4-hydroxyphenylpyruvate compared to the wild-type enzyme
N275D
-
site-directed mutagenesis
Q286E
-
site-directed mutagenesis
Q300E
-
site-directed mutagenesis
Q372E
-
site-directed mutagenesis
S260A
-
site-directed mutagenesis
E254D
site-directed mutagenesis, the mutant shows 5% remaining activity compared to the wild-type enzyme
Q375N
site-directed mutagenesis, the mutant shows that a solvent accessible channel opens to the putative substrate binding site, suggesting this is responsible for the complete loss of activity, modeling, overview. Inactive mutant
Q375N/R378K
site-directed mutagenesis, inactive mutant
R378K
site-directed mutagenesis, the mutant shows 5% remaining activity compared to the wild-type enzyme
P214T
-
site-directed mutagenesis, during hte reaction mechanism, the last 1,2 rearrangement is blocked in S246A HPPD mutant, so that an arene oxide-derived intermediate is released as an alternative product
F337I
-
0.67 of wild-type activity
F341Y
-
16.9% of wild-type activity
F364I
-
site-directed mutagenesis, the mutant enzyme produces 47% homogentisate, 15% 4-hydroxyphenylacetate, and 19% quinolacetate, which differs from the wild-type activity
N216I
-
0.15% of wild-type activity
N216I/F337I
-
inactive
N216I/F337I/F341Y
-
inactive
N216I/F341Y
-
inactive
N245D
-
site-directed mutagenesis, the mutant enzyme produces 52% homogentisate and 26.8% 4-hydroxyphenylacetate, and 21.2% quinolacetate, which differs from the wild-type activity
N245I
-
site-directed mutagenesis, the mutant enzyme produces 13% homogentisate and 87% 4-hydroxyphenylacetate, which differs from the wild-type activity
N245S
-
site-directed mutagenesis, the mutant enzyme produces 3.4% homogentisate and 6.6% 4-hydroxyphenylacetate, and 90% quinolacetate, which differs from the wild-type activity
P214T
-
1.6% of wild-type activity
P214T/F337I/F341Y
-
inactive
P214T/N216I
-
inactive
P214T/N216I/F337I
-
inactive
P214T/N216I/F337I/F341Y
-
inactive
P214T/N216I/F341Y
-
inactive
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
-
overexpression of enzyme in Nicotiana tabacum, transgenic plants have a 10-fold higher resistance to the bleaching herbicide sulcotrione, transgenic seeds have an up to two-fold enhanced level of vitamin E without change in the ratio of gamma-tocopherol and gamma-tocotrienol. Level of plastoquinone is enhanced in leaves of transgenic lines during leaf senescence
medicine
-
mechanism of binding of the inhibitor 2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione used to treat type I tyrosinemia. Enzyme-Fe(II)-inhibitor complex does not oxidize, dissociation rate constant is essentially zero
Show AA Sequence (1273 entries)
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