Information on EC 1.13.11.27 - 4-hydroxyphenylpyruvate dioxygenase

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

EC NUMBER
COMMENTARY
1.13.11.27
-
RECOMMENDED NAME
GeneOntology No.
4-hydroxyphenylpyruvate dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
homogentisate
-
-
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
rate-determining step in catalysis is a protein conformation change
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
mono-iso-ordered bi-bi mechanism with binding of 4-hydroxyphenylpyruvate before O2 and release of CO2 before homogentisate. A Theorell-Chance mechanism can not be excluded
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
enzyme form 3: ordered bi bi mechanism where 4-hydroxyphenylpyruvate is added prior to oxygen and CO2 released before homogentisate
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
evidence against participation of a quinol as a free intermediate
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
in plants this enzyme activity is involved in two distinct metabolic processes, the biosynthesis of prenylquinones and the catabolism of tyrosine
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
addition of substrate and oxygen to the holoenzyme is formally random, holo-enzym in complex with substrate has a 3600-fold increase in oxygen reactivity
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
ternary enzyme-substrate complex is firstly decarboxylated to the iron(II)-peracid intermediate followed by heterolytic cleavage of the O-O bond yielding an iron(IV)-oxospecies. This attacks the aromatic ring in C1 position. The arene oxide has no catalytic relevance
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
mechanism of oxygen binding and activation, structural relationship to other dioxygenases
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
the C-terminal helix forms a gate for substrate access to the active site around a nonheme ferrous iron center
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
the C-terminal helix forms a gate for substrate access to the active site around a nonheme ferrous iron center, completely sequestering the active site from solvent
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
The data of the rapid acid quench followd by HPLC product analysis, indicate that the homogenisate product is formed during one turnover, as the amount of product is equivalent to the enzyme-substrate complex at 191 ms.
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
reaction mechanism, overview
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
direct involvement of the arene oxide intermediate to the reaction mechanism
-
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2
show the reaction diagram
mono-iso-ordered bi-bi mechanism with binding of 4-hydroxyphenylpyruvate before O2 and release of CO2 before homogentisate. A Theorell-Chance mechanism can not be excluded
Pseudomonas sp. P.J. 874
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
decarboxylation
-
-
-
-
hydroxylation
-
-
-
-
redox reaction
-
-
-
-
side-chain migration
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Metabolic pathways
-
phenylalanine biosynthesis (cytosolic, plants)
-
phenylalanine degradation IV (mammalian, via side chain)
-
Phenylalanine metabolism
-
plastoquinol-9 biosynthesis I
-
tyrosine degradation I
-
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.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4-HPPD
-
-
-
-
4-HPPD
P32754
-
4-HPPD
-
-
4-HPPD
Q8EKK9
-
4-hydroxyphenylpyruvare dioxygenase
A7BG64
-
4-hydroxyphenylpyruvate dioxygenase
-
-
4-hydroxyphenylpyruvate dioxygenase
P80064
-
4-hydroxyphenylpyruvate dioxygenase
Q9RSJ4
-
4-hydroxyphenylpyruvate dioxygenase
Q8EKK9
-
4-hydroxyphenylpyruvate dioxygenase
Vitis vinifera x Vitis riparia, Vitis vinifera x Vitis vinifera
-
-
4-hydroxyphenylpyruvic acid dioxygenase
-
-
-
-
4HPPD
-
-
-
-
EC 1.14.2.2
-
-
-
-
EC 1.99.1.14, formerly
-
-
-
-
F Alloantigen
-
-
-
-
F protein
-
-
-
-
F-antigen homolog
-
-
-
-
HPD
-
-
-
-
HPD
Vitis vinifera x Vitis riparia, Vitis vinifera x Vitis vinifera
-
-
HPPD
-
-
-
-
HPPDase
-
-
-
-
Legiolysin
-
-
-
-
oxygenase, 4-hydroxyphenylpyruvate di-
-
-
-
-
p-hydroxyphenylpyruvate dioxygenase
-
-
-
-
p-hydroxyphenylpyruvate dioxygenase
P93836
-
p-hydroxyphenylpyruvate hydroxylase
-
-
-
-
p-hydroxyphenylpyruvate oxidase
-
-
-
-
p-hydroxyphenylpyruvic acid hydroxylase
-
-
-
-
p-hydroxyphenylpyruvic hydroxylase
-
-
-
-
p-hydroxyphenylpyruvic oxidase
-
-
-
-
T-cell reactive protein
-
-
-
-
TF-AG
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9029-72-5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
expressed in Escherichia coli
Swissprot
Manually annotated by BRENDA team
sequence of the cloned 4-hydroxyphenylpyruvate dioxygenase coincides with published 4-hydroxyphenylpyruvate dioxygenase-sequences from Arabidopsis thaliana to a nearly perfect identity match except for minor differences in the C-terminal parts of the cDNA.
Swissprot
Manually annotated by BRENDA team
Frog
-
-
-
Manually annotated by BRENDA team
enzyme form 1, 2, and 3
-
-
Manually annotated by BRENDA team
enzyme form 3
-
-
Manually annotated by BRENDA team
overexpression in Nicotiana tabacum
-
-
Manually annotated by BRENDA team
The alpha-tocophenol content is increased in leaves of the transgenic Nicotiana tabacum (tabacco) plants, whereas the transgenic tobacco seeds contain a significantly increased gamma-tocochromanol level. Overexpression of the barley 4-hydroxyphenylpyruvate dioxygenase gene in plastids of tobacco does not prove to be advantageous in comparison to transgenic tobacco plants expressing high 4-hydroxyphenylpyruvate dioxygenase level in the cytoplasma.
Swissprot
Manually annotated by BRENDA team
4-hydroxyphenylpyruvate dioxygenase of Pseudomonas fluroscens is cloned into Nicotiana tabacum (cv. PBD6) and into Glycine max (cv. Jack) and tobacco plastid transformants acquire a strong herbicide tolerance, performing better than nuclear transformants.
-
-
Manually annotated by BRENDA team
Pseudomonas sp. P.J. 874
strain P.J. 874
-
-
Manually annotated by BRENDA team
expression in Escherichia coli
SwissProt
Manually annotated by BRENDA team
strain MR-1
UniProt
Manually annotated by BRENDA team
expression in Escherichia coli
-
-
Manually annotated by BRENDA team
hog
-
-
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
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
4-hydroxyphenylpyruvate dioxygenase and hydroxymandelate synthase, HMS, EC 1.13.11.46, catalyze similar reactions using the same substrates, 4-hydroxyphenylpyruvate and dioxygen. Initially, both enzymes reduce and activate dioxygen in order to decarboxylate 4-hydroxyphenylpyruvate, yielding 4-hydroxyphenylacetate, CO2, and an activated oxo intermediate Both enzymes then hydroxylate 4-hydroxyphenylacetate but do so in different positions
evolution
P32754
the enzyme belongs to the non-haem Fe(II)/2-oxoacid-dependent oxygenase superfamily, which couples the oxidative decarboxylation of a 2-oxoacid (most commonly a-ketoglutarate) to the oxidation of the prime substrate
malfunction
P32754
deficiency in active 4-HPPD in humans results in type III tyrosinemia, a rare autosomal recessive disorder
metabolism
-
inhibition mechanism of 2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
physiological function
Vitis vinifera x Vitis riparia, Vitis vinifera x Vitis vinifera
-
involved in aromatic amino acid metabolism
metabolism
P32754
enzyme 4-HPPD catalyzes the second step in the pathway of tyrosine catabolism, the conversion of 4-hydroxyphenyl-pyruvate to homogentisate
additional information
-
substrate binding, structure modeling, structure comparisons, quantum mechanical/molecular mechanical calculations of the enzyme-substrate complex and key reaction intermediates, overview. Three different models of HPP binding within the Arabidopsis thaliana HPPD active site. Residues Ser246, Asn261, Gln272, Gln286, and Gln358 are important for catalysis
additional information
-
substrate binding, structure modeling, structure comparisons, quantum mechanical/molecular mechanical calculations of the enzyme-substrate complex and key reaction intermediates, overview. Residues Ser260, Asn275, Gln286, Gln300, and Gln372 are important for catalysis
additional information
-
substrate binding, structure modeling, structure comparisons, quantum mechanical/molecular mechanical calculations of the enzyme-substrate complex and key reaction intermediates, overview. Residues Ser201, Asn216, Gln225, Gln239, and Gln309 are important for catalysis
additional information
-
substrate binding, structure modeling, structure comparisons, quantum mechanical/molecular mechanical calculations of the enzyme-substrate complex and key reaction intermediates, overview. Residues Ser201, Asn216, Gln226, Gln230, and Gln305 are important for catalysis
additional information
P32754
the disordered C-terminal tail of the enzyme, which is extended C-terminus compared to other 4-HPPD enzymes, plays an important role in catalysis with a critical role of residue Q375 in orientating the tail and ensuring the conformation of the terminal alpha-helix to maintain the integrity of the active site for catalysis. The active site of 4-HPPD is enclosed by a C-terminal alpha-helix which is assumed to function as a gate which controls access of substrate. Interactions provided by Q375 to hold the terminal helix and the tail in proper position are critical for isolating the active site from solvent during catalysis, enzyme structure modeling, overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
Pseudomonas sp., Pseudomonas sp. P.J. 874
-
-
-
?
(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
-
-
-
-
?
3,4-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
homogentisate + CO2
show the reaction diagram
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Q9RSJ4, -
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
A7BG64
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Q8EKK9, -
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
P32754
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Q8EKK9, -
-
homogentisate is an intermediate of the tyrosine degradation pathway, and a precursor of a redox-cycling metabolite, pyomelanin
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
best substrate
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
activity with the keto-form of 4-hydroxyphenylpyruvate is 40times higher than with the enol tautomer
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
evidence against participation of a quinol as a free intermediate
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
enzyme participates in catabolism of tyrosine
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
key enzyme involved in tyrosine catabolism, congenital 4-hydroxyphenylpyruvate dioxygenase deficiency is a rare, relatively benign condition known as hereditary type III tyrosinemia
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
enzyme is involved in production of homogentisate, the aromatic precursor of all phenylquinones
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
4-hydroxyphenylpyruvate dioxygenase does not use phenolpyruvate as a substrate in normal plant metabolism
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
hppD is transcriptionally activated by HpdA and repressed by HpdR
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
P32754
the conversion of 4-hydroxyphenyl-pyruvate to homogentisate involves oxidative decarboxylation, side-chain migration and aromatic hydroxylation in a single catalytic cycle. The 4-HPPD reaction is unusual in that the 2-oxoacid and prime substrate moieties are contained within the same molecule
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Pseudomonas sp. P.J. 874
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Pseudomonas sp. P.J. 874
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Pseudomonas sp. P.J. 874
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
2,5-dihydroxyphenylacetate + CO2
show the reaction diagram
P80064
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2 + 4-hydroxyphenylacetate
show the reaction diagram
-
-
the wild-type enzyme produces 85% homogentisate and 15% 4-hydroxyphenylacetate
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2 + 4-hydroxyphenylacetate
show the reaction diagram
-
-
the wild-type enzyme produces 90% homogentisate, 1% quinolacetate, and 9% 4-hydroxyphenylacetate
-
?
4-hydroxytetrafluorophenylpyruvate + O2
? + CO2
show the reaction diagram
-
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
-
-
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
-
at 10% of the activity with 4-hydroxyphenylpyruvate
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
Pseudomonas sp. P.J. 874
-
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
Pseudomonas sp. P.J. 874
-
-
-
?
phenylpyruvate + O2
2-hydroxyphenylacetate + CO2
show the reaction diagram
Pseudomonas sp. P.J. 874
-
-
-
-
?
[(4-hydroxyphenyl)thio]pyruvate + O2
[(4-hydroxyphenyl)sulfinyl]acetate + CO2
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. P.J. 874
-
substrate undergoes oxidative decarboxylation and sulfoxidation to give [(4-hydroxyphenyl)sulfinyl]acetate, ring oxidation is not observed
-
-
?
4-methylphenylpyruvate + O2
? + CO2
show the reaction diagram
-
poor substrate
-
-
?
additional information
?
-
-
Single turnover kinetics are observed spectrophotometrically by mixing equal volumes of the anaerobic enzyme-substrate complex and solutions containing molecular oxygen in the presence of saturating HPP using a stopped-flow spectrophotometer. k1 = 7.4 x 100000 /M/s, k2 = 74 /s, k3 = 13.2 /s, k4 = 1.6 /s, indicating the accumulation of three catalytic intermediates. The kinetic data observed with substrate substituted with deutererons for aromatic protons are not significantly different from those observed with the proteo substrate. The final phase in catalysis (k4), decreases to 0.7 /s in the presence of deuterium oxide solvent, indicating the involvement of a solvent-derived proton in this step.
-
-
-
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
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Q9RSJ4, -
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
A7BG64
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
Q8EKK9, -
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
enzyme participates in catabolism of tyrosine
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
key enzyme involved in tyrosine catabolism, congenital 4-hydroxyphenylpyruvate dioxygenase deficiency is a rare, relatively benign condition known as hereditary type III tyrosinemia
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
enzyme is involved in production of homogentisate, the aromatic precursor of all phenylquinones
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
-
hppD is transcriptionally activated by HpdA and repressed by HpdR
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2
show the reaction diagram
P32754
the conversion of 4-hydroxyphenyl-pyruvate to homogentisate involves oxidative decarboxylation, side-chain migration and aromatic hydroxylation in a single catalytic cycle. The 4-HPPD reaction is unusual in that the 2-oxoacid and prime substrate moieties are contained within the same molecule
-
-
?
4-hydroxyphenylpyruvate + O2
2,5-dihydroxyphenylacetate + CO2
show the reaction diagram
P80064
-
-
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2 + 4-hydroxyphenylacetate
show the reaction diagram
-
-
the wild-type enzyme produces 85% homogentisate and 15% 4-hydroxyphenylacetate
-
?
4-hydroxyphenylpyruvate + O2
homogentisate + CO2 + 4-hydroxyphenylacetate
show the reaction diagram
-
-
the wild-type enzyme produces 90% homogentisate, 1% quinolacetate, and 9% 4-hydroxyphenylacetate
-
?
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 ACCESSION NO.
COMMENTARY
LITERATURE
copper
-
0.4 atoms of Fe per 89000 Da enzyme. Removal of copper does not correlate with loss of activity
copper
-
variable amounts of copper: 0.2-0.5 mol per mol of enzyme
Fe2+
-
restores activity after inactivation by Fe2+-chelators
Fe2+
-
essential role for enzyme-bound Fe2+ in catalytic process
Fe2+
-
the enol tautomer of 4-hydroxyphenylpyruvate appears to form a 2:1 complex with enzyme-bound Fe2+, the tautomerizable keto group with a double bond in 3-4 position favors productive substrate binding to Fe2+
Fe2+
-
essential role for enzyme bound zinc
Fe2+
-
in octahedral complex with His205, His287, Glu373 and three water molecules
Fe2+
-
required for activity, the active-site iron is in the ferric oxidation state and coordinated by the side chains of two histidines and on glutamate
Fe2+
-
required for activity
Fe2+
Q9RSJ4, -
-
Fe2+
-
a Fe(II)-dependent dioxygenase, Fe2+ is involved in the catalytic mechanism binding to the substrate, overview
Fe3+
-
iron in the Fe3+ form must be reduced for enzyme activity
Fe3+
-
ferric iron in a rhombic ligand field
Iron
-
0.88 mol of iron per mol of enzyme
Iron
-
iron-tyrosinate protein, high spin ferric center in a rhombic environment
Iron
-
0.95 mol of iron per mol of enzyme
Iron
-
0.9 atoms of Fe per 89000 Da enzyme. Removal of iron correlates with loss of activity
Iron
-
variable amounts of iron: 0.6-1.3 mol per mol of enzyme
Iron
-
contains 1 atom of iron per enzyme subunit
Iron
-
one non-heme iron atom per monomer liganded to the sidechains of His161, His240, Glu322 and one acetate molecule
Iron
-
distorted square-pyramidal coordination with Glu349, His187, His270
Iron
-
nonheme ferrous iron
Zinc
-
0.68 mol of zinc per mol of enzyme
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(2,6-difluoro-4-hydroxyphenyl)pyruvate
-
competitive, most potent
1,10-phenanthroline
-
-
1,10-phenanthroline
-
0.01 mM, 50% inhibition
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
-
50% inhibition at 7 nM
1-(1,1-dimethylethyl)-5-hydroxy-4-(3-(4-(methoxy)phenyl)-2-methyl-4-(methylsulfonyl)benzoyl)pyrazole
P32755
50% inhibition below 20 nM
1-(2'-theonyl)-2,3-trifluoroacetone
-
0.0049 mM, 50% inhibition
-
1-(2'-thionyl)-3,3,3-trifluoroacetone
-
-
2,2'-Bipyridyl
-
-
2,2'-Bipyridyl
-
1.8 mM, 50% inhibition
2,2'-Bipyridyl
-
0.32 mM, 50% inhibition
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
Q8EKK9, -
competitive inhibitor
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione
-
potent linear competitive inhibitor
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione
-
-
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione
-
-
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione
-
rapid inactivation of the enzyme by the formation of an enzyme-inhibitor complex that dissociates with recovery of enzyme activity
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
-
product inhibition
2-hydroxyphenylacetate
-
10 mM, 50% inhibition, noncompetitive
2-hydroxyphenylacetate
-
-
2-Hydroxyphenylpyruvate
-
competitive
2-thienylpyruvate
-
mechanism-based inhibitor
2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
-
mechanism of binding, enzyme-Fe(II)-inhibitor complex does not oxidize, dissociation rate constant is essentially zero
2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
-
-
2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
-
is a selective and efficient herbicide that can be applied as a medicine in a hereditary metabolic disease - tyrosinemia type I
2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
P80064
herbicide, drug to treat inborn defects of tyrosine catabolism, specifically type I tyrosinemia and alkaptonuria
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
A7BG64
-
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
-
above 0.2 mM; substrate inhibition
4-hydroxyphenylpyruvate
-
substrate inhibition
4-hydroxyphenylpyruvate
-
interaction of enol-4-hydroxyphenylpyruvate with enzyme-bound Fe3+, formed by autooxidation, causes the substrate inhibition
4-hydroxyphenylpyruvate
-
enzyme form 2 and 3, no substrate inhibition is observed for enzyme form 1
4-hydroxyphenylpyruvate
-
-
4-hydroxyphenylpyruvate
-
above 0.2 mM
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
Bathocuproine
-
3 mM, 50% inhibition
bathophenanthroline
-
-
bathophenanthroline
-
-
bathophenanthroline
-
0.018 mM, 50% inhibition; reactivation to 30% by dialysis and to 60% by addition to Fe2+
bathophenanthroline
-
-
bathophenanthroline
-
0.012 mM, 50% inhibition
benzobicyclon hydrolysate
A7BG64
-
Cupferron
-
-
Cupferron
-
0.0011 mM, 50% inhibition
Cupferron
-
0.0071 mM, 50% inhibition
diethyl dicarbonate
-
5 mM, 50% inhibition
diethyldithiocarbamate
-
-
diethyldithiocarbamate
-
0.0049 mM, 50% inhibition; restored by dialysis
diethyldithiocarbamate
-
-
diethyldithiocarbamate
-
0.0021 mM, 50% inhibition
diketonitrile of isoxaflutole
-
half-site reactivity, nearly irreversible
Dithizone
-
-
Dithizone
-
0.14 mM, 50% inhibition
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
ethanol
-
5% v/v, 50% inhibition, complete loss of activity at 20% v/v
Fe2+
-
1 mM, 30% inhibition
-
Fe2+-chelators
-
-
-
grandiflorone
P93836
-
H2O2
-
0.01 mM-1 mM, 5-15% inhibition
H2O2
-
10 nM, 60% inhibition. 0.001 mM, complete inhibition
H2O2
-
10 mM, 30% inhibition
Homogentisate
-
product inhibition
Homogentisate
-
-
L-Phe
-
10 mM, 50% inhibition, competitive
leptospermone
P93836
-
LnCl3
-
1 mM, 50% inhibition
-
manuka oil
P93836
-
-
o-hydroxyphenylpyruvate
-
competitive inhibitor of 4-hydroxyphenylpyruvate
oxaloacetate
-
10 mM, 50% inhibition, noncompetitive
pentafluorophenylpyruvate
-
competitive
phenylglyoxalate
-
50% inhibition above 10 mM, competitive
phenylpyruvate
-
non-competitive inhibition with 4-hydroxyphenylpyruvate as substrate
phenylpyruvate
-
0.53 mM, 50% inhibition, competitive
phenylpyruvate
-
competitive
pyridoxal 5'-phosphate
-
2 mM, 50% inhibition
pyruvate
-
50% inhibition above 10 mM, competitive
sulcotrione
P93836
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
P93836
-
-
YCl3
-
1 mM, 50% 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
mesotrione
-
binding constant of 602 microM
additional information
-
neither epoxybenzoaquinone ester nor 2-hydroxy-3-phenyl-3-butenoic acid is an inhibitor
-
additional information
-
the enzyme is molecular target of new families of potent herbicides
-
additional information
P32755
[1-tert-butyl-3-(2,4-dichlorophenyl)-5-hydroxy-1H-pyrazol-4-yl][2-chloro-4-(methylsulfonyl)phenyl]methanone, no significant inhibition up to 0.02 mM
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1-butanol
-
5% v/v, 2.9fold activation
1-Propanol
-
10% v/v, 2.1fold activation
2,6-dichlorophenol indophenol
-
reducing cofactor required, either ascorbate, or a combination of 2,6-dichlorophenol indophenol and glutathione
2,6-dichlorophenol indophenol
-
competitive activator
2,6-dichlorophenol indophenol
-
stimulates
acetone
-
10% v/v, 5.7fold activation
acetonitrile
-
10% v/v, 3.6fold activation
ascorbate
-
reducing cofactor required, either ascorbate, or a combination of 2,6-dichlorophenol indophenol and glutathione in presence of catalase
ascorbate
-
an optimized combination of ascorbate, 50 mM, catalase, 2.4 g/l, and iron, 0.05 mM, activates more than a combination of 2,6-dichlorophenolindophenol, glutathione and catalase
ascorbate
-
required for activity
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
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
-
methyl-cellosolve
-
10% v/v, 2.2fold activation
additional information
P93836
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]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
4-hydroxyphenylpyruvate
-
-
0.0069
-
4-hydroxyphenylpyruvate
-
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
0.0075
-
4-hydroxyphenylpyruvate
-
recombinant wild-type enzyme, pH and temperature not specified in the publication
0.0087
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant P243T
0.009
-
4-hydroxyphenylpyruvate
-
recombinant mutant S246A, pH and temperature not specified in the publication
0.015
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant N245Q
0.025
-
4-hydroxyphenylpyruvate
-
enzyme form 1
0.027
-
4-hydroxyphenylpyruvate
-
enzyme form 2
0.027
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, wild-type enzyme and mutant N245I
0.029
-
4-hydroxyphenylpyruvate
-
enzyme form 3
0.03
-
4-hydroxyphenylpyruvate
-
-
0.03
-
4-hydroxyphenylpyruvate
-
-
0.04
-
4-hydroxyphenylpyruvate
-
-
0.04
-
4-hydroxyphenylpyruvate
-
-
0.041
-
4-hydroxyphenylpyruvate
-
-
0.0432
-
4-hydroxyphenylpyruvate
A7BG64
-
0.047
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant S230A
0.05
-
4-hydroxyphenylpyruvate
-
-
0.07
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAL381
0.075
-
4-hydroxyphenylpyruvate
-
-
0.08
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, wild-type enzyme and mutant DELTAG388
0.09
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAL385
0.11
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant E254D
0.12
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAG379
0.153
-
4-hydroxyphenylpyruvate
-
recombinant mutant N261D, pH and temperature not specified in the publication
0.24
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant R378K
0.286
-
4-hydroxyphenylpyruvate
-
recombinant mutant Q358E, pH and temperature not specified in the publication
0.3
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAN380
0.452
-
4-hydroxyphenylpyruvate
-
recombinant mutant Q272E, pH and temperature not specified in the publication
0.5
-
4-hydroxyphenylpyruvate
Frog
-
-
0.551
-
4-hydroxyphenylpyruvate
-
recombinant mutant Q286E, pH and temperature not specified in the publication
11
-
4-hydroxyphenylpyruvate
-
Solvent is H2O. Steady-state analyses of 4-hydroxyphenylpyruvate dioxygenase using proteo-HPP, 2,3,5,6-tetradeutero-HPP, and proteo-HPP in deuterium oxide are carried out in 1 ml assays on a Hasetech oxygen electrode. The assay mixture include ferrous sulfate (10 microM), DTT (1mM), HPPD (500 nM), and HPP (0-1 mM) in 20 mM HEPES, pH (pD) 7.0, under conditions of atmospheric oxygen (350 microM at 5C)
22
-
4-hydroxyphenylpyruvate
-
Solvent is H2O. Steady-state analyses of 4-hydroxyphenylpyruvate dioxygenase using proteo-HPP, 2,3,5,6-tetradeutero-HPP, and proteo-HPP in deuterium oxide are carried out in 1 ml assays on a Hasetech oxygen electrode. The assay mixture include ferrous sulfate (10 microM), DTT (1mM), HPPD (500 nM), and HPP (0-1 mM) in 20 mM HEPES, pH (pD) 7.0, under conditions of atmospheric oxygen (350 microM at 5C)
54
-
4-hydroxyphenylpyruvate
-
Solvent is D2O. Steady-state analyses of 4-hydroxyphenylpyruvate dioxygenase using proteo-HPP, 2,3,5,6-tetradeutero-HPP, and proteo-HPP in deuterium oxide are carried out in 1 ml assays on a Hasetech oxygen electrode. The assay mixture include ferrous sulfate (10 microM), DTT (1mM), HPPD (500 nM), and HPP (0-1 mM) in 20 mM HEPES, pH (pD) 7.0, under conditions of atmospheric oxygen (350 microM at 5C)
0.069
-
O2
-
pH 7.0, 25C
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
-
phenylpyruvate
-
-
0.066
-
phenylpyruvate
-
-
0.5
-
phenylpyruvate
-
-
0.5
-
phenylpyruvate
-
-
0.52
-
phenylpyruvate
-
-
0.05
-
4-hydroxytetrafluorophenylpyruvate
-
-
additional information
-
additional information
-
the Km-value is the same at 25C and at 38C
-
additional information
-
additional information
-
-
-
additional information
-
additional information
-
-
-
additional information
-
additional information
-
the Km-value is the same at 25C and at 38C
-
additional information
-
additional information
-
Kd value for 4-hydroxyphenylpyruvate with 4-hydroxyphenylpyruvate dioxygenase Fe2+ is 29 microM, whereas the binding constant for phenylpyruvate is determined to be 319 microM
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
6.8
-
(4-hydroxyphenyl)-pyruvate
-
-
0.03
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant N245Q
0.06
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant E254D
0.065
-
4-hydroxyphenylpyruvate
-
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
0.07
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant R378K
0.1
-
4-hydroxyphenylpyruvate
-
recombinant mutant Q286E, pH and temperature not specified in the publication
0.1
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAG379
0.2
-
4-hydroxyphenylpyruvate
-
recombinant mutant Q272E, pH and temperature not specified in the publication; recombinant mutant S246A, pH and temperature not specified in the publication
0.3
-
4-hydroxyphenylpyruvate
-
recombinant mutant N261D, pH and temperature not specified in the publication
0.5
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAN380
0.64
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant N245I
1
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAL381
1.4
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAG388 and DELTAL385
1.8
-
4-hydroxyphenylpyruvate
-
recombinant wild-type enzyme, pH and temperature not specified in the publication
2.1
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant P243T
2.1
-
4-hydroxyphenylpyruvate
-
recombinant mutant Q358E, pH and temperature not specified in the publication
2.2
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, wild-type enzyme
2.8
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant S230A
6.8
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, wild-type enzyme
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.27
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant R378K
4595
0.57
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant E254D
4595
0.9
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAG379
4595
1.5
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAN380
4595
2.4
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant N245Q
4595
14
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAL381
4595
16
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAL385
4595
17
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, mutant DELTAG388
4595
24
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant N245I
4595
29
-
4-hydroxyphenylpyruvate
P32754
pH 6.5, 37C, wild-type enzyme
4595
60
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant S230A
4595
241
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, mutant P243T
4595
254
-
4-hydroxyphenylpyruvate
-
pH 7.0, 25C, wild-type enzyme
4595
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0013
-
(2,6-difluoro-4-hydroxyphenyl)pyruvate
-
-
0.0000098
-
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexanedione
-
-
0.4
-
4-hydroxyphenylpyruvate
-
about, enzyme form 3
0.014
-
pentafluorophenylpyruvate
-
-
0.15
-
thiophenol oxalate
-
-
0.7
-
4-hydroxyphenylpyruvate
-
about, enzyme form 2
additional information
-
additional information
-
-
-
additional information
-
additional information
-
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00675
-
4-(2,4-dichlorobenzyl)-1,3-dimethyl-5-hydroxypyrazole
A7BG64
recombinant enzyme
0.00017
-
4-(3-cyclohexylpropanoyl)-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
0.00227
-
4-(3-cyclohexylpropanoyl)-5-hydroxy-2,2,6-trimethyl-6-(3-methylbut-2-enyl)cyclohex-4-ene-1,3-dione
-
-
0.00025
-
4-dodecanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
0.00096
-
4-hexanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
0.0143
-
5-hydroxy-2,2,6,6-tetramethyl-4-(2-methylbutanoyl)cyclohex-4-ene-1,3-dione
-
-
0.0189
-
5-hydroxy-2,2,6,6-tetramethyl-4-(2-phenylacetyl)cyclohex-4-ene-1,3-dione
-
-
0.0118
-
5-hydroxy-2,2,6,6-tetramethyl-4-(3-methylbutanoyl)cyclohex-4-ene-1,3-dione
-
-
0.00075
-
5-hydroxy-2,2,6,6-tetramethyl-4-(3-phenylpropanoyl)cyclohex-4-ene-1,3-dione
-
grandiflorone, most active naturally occurring beta-triketone
0.0445
-
5-hydroxy-4-isobutyryl-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
0.000015
-
topramezone
-
enzyme isolated from Setaria faberi
0.000023
-
topramezone
-
enzyme recombinantly expressed in Arabidopsis thaliana
additional information
-
3,3,5,5-tetramethyl-2,4,6-trioxocyclohexyl 3-methylbutanoate
-
IC50 value above 50 microM
additional information
-
3-(2-cyclohexylethyl)-5,5,7,7-tetramethylbenzo[d]isoxazole-4,6(5H,7H)-dione
-
IC50 value above 50 microM
additional information
-
3-cyclohexyl-1-(2,4,6-trihydroxy-3-(3-methylbut-2-enyl)phenyl)propan-1-one
-
IC50 value above 50 microM
0.00041
-
4-(3-cyclohexylpropanoyl)-5-methoxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
additional information
-
4-(cyclohexanecarbonyl)-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
IC50 value above 50 microM
additional information
-
4-acetyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
IC50 value above 50 microM
0.000019
-
4-decanoyl-5-hydroxy-2,2,6,6-tetramethylcyclohex-4-ene-1,3-dione
-
-
additional information
-
4-dodecanoyl-5-hydroxy-2,2,6,6-tetrakis(3-methylbut-2-enyl)cyclohex-4-ene-1,3-dione
-
IC50 value above 50 microM
0.00031
-
5-hydroxy-2,2,6,6-tetramethyl-4-palmitoylcyclohex-4-ene-1,3-dione
-
-
additional information
-
5-hydroxy-4-isobutyryl-2,2,6,6-tetrakis(3-methylbut-2-enyl)cyclohex-4-ene-1,3-dione
-
IC50 value above 50 microM
0.00234
-
benzobicyclon hydrolysate
A7BG64
recombinant enzyme
additional information
-
additional information
P93836
IC50 values: 0.015 mg/ml for manuka oil, 0.00402 mg/ml for triketone-rich fraction, 0.00314 mg/ml for leptospermone and 0.00022 mg/ml for grandiflorone
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.004
-
P93836
protein expressed in Escherichia coli DH5alpha
0.04
-
P32754
purified recombinant mutant R378K, pH 6.5, 37C, homogentisate production
0.06
-
P32754
purified recombinant mutant E254D, pH 6.5, 37C, O2 consumption
0.07
-
P32754
purified recombinant mutant R378K, pH 6.5, 37C, O2 consumption
0.11
0.2
P93836
protein expressed in Escherichia coli SG13009
0.17
0.26
P93836
protein expressed in Escherichia coli M15(pRep4)
0.2
-
P32754
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
-
-
0.92
-
-
-
2.33
-
-
-
2.6
-
P32754
purified recombinant wild-type enzyme, pH 6.5, 37C, O2 consumption
2.8
-
P32754
purified recombinant wild-type enzyme, pH 6.5, 37C, homogentisate production
10.6
-
-
pH 7.0, 25C
39.6
-
-
-
51
-
-
-
additional information
-
-
the oxygen electrode provides an excellent measure of enzyme activity under conditions where maximum enzyme activity is obtainable in the absence of a reducing agent
additional information
-
-
methods for assay
additional information
-
-
with substrate 2,3,5,6-tetraproteo 4-hydroxyphenylpyruvate and the solvent H2O Vmax = 1.9/s, with substrate 2,3,5,6-tetradeutero 4-hydroxyphenylpyruvate and the solvent H2O Vmax = 1.8/s, with substrate 2,3,5,6-tetraproteo 4-hydroxyphenylpyruvate and the solvent D2O Vmax = 0.86/s
additional information
-
P93836
activity of 0.0033 micromol/min/mg for 2YT-grown Escherichia coli DH5alpha extract and 0.0018 micromol/min/mg for LB-grown Escherichia coli DH5alpha
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
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
6.5
-
P32754
assay at
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
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
P32754
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
enzymatic rate increases approximately 5fold for every 10C increase, Km-value is the same at 25C and at 38C
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.84
-
Q9RSJ4, -
predicted
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
P32755
abundantly expressed in neurons of
Manually annotated by BRENDA team
P32755
abundantly expressed in neurons of
Manually annotated by BRENDA team
-
photosynthetically active tissue
Manually annotated by BRENDA team
P93836
drought-stressed Arabidopsis thaliana leaf RNA
Manually annotated by BRENDA team
-
etiolated
Manually annotated by BRENDA team
additional information
-
no enzymatic activity in any tissue other than liver and kidney
Manually annotated by BRENDA team
additional information
-
-
Manually annotated by BRENDA team
additional information
-
no activity detected in heart, muscle and brain
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
40000
-
-
in transgenic Glycine max (cv. Jack), SDS-PAGE; in transgenic Nicotiana tabacum (cv. PBD6), Western blot
50000
54000
-
gel filtration
52540
-
Q9RSJ4, -
predicted molecular mass
63000
67000
-
enzyme forms 1, 2, and 3, gel filtration
66000
-
-
gel filtration
85000
-
-
gel filtration
87000
-
-
equilibrium sedimentation
89000
-
-
method not mentioned
89000
-
-
equilibrium ultracentrifugation
97000
-
-
equilibrium sedimentation
100000
-
-
non-denaturing PAGE
150000
-
-
high-speed equilibrium sedimentation
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 48094, calculation from nucleotide sequence
?
-
x * 48000, SDS-PAGE
?
P32755
x * 44000, SDS-PAGE and deduced from gene sequence
dimer
-
2 * 44000, nonidentical subunits, SDS-PAGE
dimer
-
2 * 43000, SDS-PAGE
dimer
-
2 * 49000, SDS-PAGE
dimer
-
weakly associated, crystallization data
dimer
-
crystallization data
homodimer
-
crystallization data
homodimer
P32754
2 * 45000, SDS-PAGE
tetramer
-
4 * 36000, SDS-PAGE
tetramer
Pseudomonas sp. P.J. 874
-
4 * 36000, SDS-PAGE; 4 * 36000, SDS-PAGE
-
additional information
-
the multiple enzyme forms are not interconvertible by oxidation, reduction, pyridine inactivation, metal chelator treatment, or treatment with neuraminidase
additional information
-
conditions which are known to accelerate nonenzymic deamination of proteins cause interconversion of multiple molecular forms
additional information
P32754
five residues of the N-terminus in recombinant 4-HPPD are TTYSD
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
phosphoprotein
-
recombinant enzyme can be phosphorylated in human AMA cells, phosphorylation is not indispensable for activity
proteolytic modification
P32755
undergoes both N-and C-terminal processing, C-terminal processing enhances enzymic activity
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
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
P32755
Fe(II)-form in complex with inhibitor 2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione
-
substrate-free form
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.5
10
-
37C, 30 min, in presence of Fe2+, stable
5.5
8
-
37C, 15 min, stable
7.5
-
-
37C, 7.5 min, stable
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
-20
-
P93836
enzyme activity is stable for a few weeks.
37
-
-
pH 5.5-8, 15 min, stable
37
-
-
pH 7.5, 0.64 M Tris-HCl, stable for 7.5 min
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
regeneration of time-dependent inactivated 4-hydroxyphenylpyruvate dioxygenase by incubation with ascorbic acid and/or Fe2+ salts is not successful
P93836
the enzyme is stable for several hours at room temperature, indefinitely at 4C
-
long-term storage of concentrated solutions in presence of air leads to polymerization
-
stable to freezing and thawing
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
routinely stored in presence of O2
-
395377
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
storage at room temperature results in residual enzyme activities of ca. 60-80% after 16 h or 40-50% after 30 h.
P93836
-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
-
-60C, 0.2 M NaCl solution buffered with 10 mM potassium phosphate, pH 6.7, stable for several months
-
-60C, stable for several months
-
-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
-
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
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
immobilized metal affinity chromatography
-
Q Sepharose anion exchange column, Sephacryl S-200 gel filtration
-
recombinant enzyme expressed in Escherichia coli
-
recombinant enzyme 15fold from Escherichia coli by anion-exchange and hydrophobic interaction chromatography, and gel filtration
P32754
recombinant enzyme expressed in Escherichia coli
-
at 5C or below
-
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 ACCESSION NO.
LITERATURE
overproduction of the recombinant enzyme in Escherichia coli JM105, overexpression of the complete coding sequence in transgenic Nicotiana tabacum using Agrobacterium tumefaciens transformation
-
expressed in Escherichia coli BL21 DE3
-
expressed in Eschrichia coli stains DH5alpha, M15(pRep4) or SG13009
P93836
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
-
expression in Escherichia coli
A7BG64
expression in Escherichia coli
-
DNA and amino acid sequence determination and analysis, human enzyme expression in Escherichia coli
P32754
vaccinia virus-based expression in human AMA cells and expression in Escherichia coli
-
barley 4-hydroxyphenylpyruvate dioxygenase gene introduced into the plastome of Nicotiana tabacum (tobacco)
-, O48604
expressed in Glycine max (cv. Jack); expressed in Nicotiana tabacum (cv. PBD6)
-
expression in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
recombinant expression of wild-type and mutant enzymes
-
expression in Escherichia coli
Q09PG8, -
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
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
Q9RSJ4, -
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
Q9RSJ4, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
Q375N
P32754
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
P32754
site-directed mutagenesis, inactive mutant
R378K
P32754
site-directed mutagenesis, the mutant shows 5% remaining activity compared to the wild-type enzyme
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
N245Q
-
site-directed mutagenesis, the mutant enzyme produces 44.5% homogentisate and 52% 4-hydroxyphenylacetate, and 4% quinolacetate, which differs from the wild-type activity
N245Q
-
site-directed mutagenesis, the mutant enzyme produces 52% homogentisate and 45% 4-hydroxyphenylacetate, and 3% quinolacetate, 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
P243T
-
site-directed mutagenesis, the mutant enzyme produces 13% homogentisate, 8.7% 4-hydroxyphenylacetate, and 78% quinolacetate, which differs from the wild-type activity
P243T
-
site-directed mutagenesis, the mutant enzyme produces 8.4% homogentisate, 46.5% 4-hydroxyphenylacetate, and 45% quinolacetate, which differs from the wild-type activity
S230A
-
site-directed mutagenesis, the mutant enzyme produces 11.6% homogentisate, 30.6% 4-hydroxyphenylacetate, and 57.8% quinolacetate, which differs from the wild-type activity
S230A
-
site-directed mutagenesis, the mutant enzyme produces 7% homogentisate, 57% 4-hydroxyphenylacetate, and 36% quinolacetate, which differs from the wild-type activity
E254D
P32754
site-directed mutagenesis, the mutant shows 5% remaining activity compared to the wild-type enzyme
additional information
P32754
human 4-HPPD possesses an extended C-terminus compared to other 4-HPPD enzymes. Successive truncation of the disordered tail which follows the final alpha-helix results in no changes in the Km value for 4-HPP substrate but the kcat values are significantly reduced by ca. 25 to 50%. The different conformation of E254, R378 and Q375 in the final helix might be the cause of the activity loss
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
additional information
Q8EKK9, -
construction and complementation of melA mutants of Shewanella oneidensis MR-1, in-frame deletion mutagenesis
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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