Information on EC 1.13.11.6 - 3-hydroxyanthranilate 3,4-dioxygenase

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

EC NUMBER
COMMENTARY
1.13.11.6
-
RECOMMENDED NAME
GeneOntology No.
3-hydroxyanthranilate 3,4-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
3-hydroxyanthranilate + O2 = 2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
mechanism
-
3-hydroxyanthranilate + O2 = 2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
reaction mechanism, overview
-
3-hydroxyanthranilate + O2 = 2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
2-nitrobenzoate degradation I
-
Metabolic pathways
-
tryptophan degradation to 2-amino-3-carboxymuconate semialdehyde
-
tryptophan degradation XI (mammalian, via kynurenine)
-
tryptophan degradation XII (Geobacillus)
-
Tryptophan metabolism
-
SYSTEMATIC NAME
IUBMB Comments
3-hydroxyanthranilate:oxygen 3,4-oxidoreductase (decyclizing)
Requires Fe2+.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3-HAD
Cavia porcellus Hartley
-
-
-
3-HAO
Rattus norvegicus Wistar
-
-
-
3-hydroxyanthranilate 3,4-dioxygenase
Q0VCA8
-
3-hydroxyanthranilate 3,4-dioxygenase
-
-
3-hydroxyanthranilate oxygenase
-
-
-
-
3-hydroxyanthranilic acid oxidase
-
-
-
-
3-hydroxyanthranilic acid oxygenase
-
-
-
-
3-hydroxyanthranilic acid oxygenase
-
-
3-hydroxyanthranilic acid oxygenase
Rattus norvegicus Wistar
-
-
-
3-hydroxyanthranilic oxygenase
-
-
-
-
3HAO
-
-
-
-
3HAO
Q0VCA8
-
EC 1.13.1.6
-
-
formerly
-
oxygenase, 3-hydroxyanthranilate 3,4-di-
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9029-50-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Hartley
-
-
Manually annotated by BRENDA team
Cavia porcellus Hartley
Hartley
-
-
Manually annotated by BRENDA team
recombinantly expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
rainbow trout
-
-
Manually annotated by BRENDA team
baboon
-
-
Manually annotated by BRENDA team
strain KU-7
SwissProt
Manually annotated by BRENDA team
Pseudomonas fluorescens KU-7
strain KU-7
SwissProt
Manually annotated by BRENDA team
Wistar strain
-
-
Manually annotated by BRENDA team
Rattus norvegicus Wistar
Wistar strain
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
the enzyme catalyzes the degradation of 3-hydroxyanthranilate to quinolinate in the presence of dioxygen
evolution
-
HAD belongs to the class of nonheme iron(II) enzymes and shares functional similarity with extradiol-cleaving catechol dioxygenases
additional information
-
oxidative CC bond cleavage of 2-amino-4-tert-butylphenolate on complex nonheme iron(II) complex, [(6-Me3-TPA)FeII(4-tBu-HAP)](ClO4) is mimicking the enzyme function. The iron(II)-aminophenolate complex reacts with molecular oxygen in acetonitrile under ambient conditions, 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-amino-3-hydroxybenzoic acid + O2
2,3-pyridinedicarboxylic acid (quinolinic acid) + H2O
show the reaction diagram
Q0VCA8
-
intermediate 2-amino-3-carboxymuconic acid semialdehyde
-
?
3-hydroxy-4-methylanthranilic acid + O2
?
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
r
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-, Q83V26
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
P47096, -
-
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
the enzymatic product subsequently cyclizes to quinolinate
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
high selectivity for substrate
reacts spontaneously to quinolinic acid
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
in mammalian peripheral organs the enzyme constitutes a link in the catabolic pathway of tryptophan to NAD
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
enzyme occurs in the metabolic pathway of the conversion of tryptophan to nicotinic acid
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
biosynthetic enzyme of the endogenous excitotoxin quinolinic acid
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
in the kynurenine pathway
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
Pseudomonas fluorescens KU-7
Q83V26
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
Rattus norvegicus Wistar
-
-
-
-
?
4-ethyl-3-hydroxyanthranilate + O2
2-amino-3-carboxy-4-ethylmuconate semialdehyde
show the reaction diagram
-
-
-
?
4-methyl-3-hydroxyanthranilate + O2
2-amino-3-carboxy-4-methylmuconate semialdehyde
show the reaction diagram
-
-
-
?
4-propyl-3-hydroxyanthranilic acid + O2
2-amino-3-carboxy-4-propylmuconate semialdehyde
show the reaction diagram
-
-
-
?
additional information
?
-
-, Q83V26
no detectable activity with 3-amino-4-hydroxybenzoic acid, 4-aminoresorcinol, 2-amino-m-cresol, 4-amino-3-hydroxybenzoic acid, 3-aminosalicylic acid, 6-amino-m-cresol, 3-methylcatechol, 4-methylcatechol, 1,2,4-trihydroxybenzene, 2,3-dihydroxybenzoic acid, 4-amino-m-cresol, 5-aminosalicylic acid, gentisic acid, homogentisic acid, 2-amino-4-chlorophenol, 2-amino-p-cresol, catechol, 1,2,3-trihydroxybenzene, protocatechuic acid, hydroquinone as substrates
-
-
-
additional information
?
-
Pseudomonas fluorescens KU-7
Q83V26
no detectable activity with 3-amino-4-hydroxybenzoic acid, 4-aminoresorcinol, 2-amino-m-cresol, 4-amino-3-hydroxybenzoic acid, 3-aminosalicylic acid, 6-amino-m-cresol, 3-methylcatechol, 4-methylcatechol, 1,2,4-trihydroxybenzene, 2,3-dihydroxybenzoic acid, 4-amino-m-cresol, 5-aminosalicylic acid, gentisic acid, homogentisic acid, 2-amino-4-chlorophenol, 2-amino-p-cresol, catechol, 1,2,3-trihydroxybenzene, protocatechuic acid, hydroquinone as substrates
-
-
-
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
2-amino-3-hydroxybenzoic acid + O2
2,3-pyridinedicarboxylic acid (quinolinic acid) + H2O
show the reaction diagram
Q0VCA8
-
intermediate 2-amino-3-carboxymuconic acid semialdehyde
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-, Q83V26
-
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
in mammalian peripheral organs the enzyme constitutes a link in the catabolic pathway of tryptophan to NAD
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
enzyme occurs in the metabolic pathway of the conversion of tryptophan to nicotinic acid
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
biosynthetic enzyme of the endogenous excitotoxin quinolinic acid
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
-
in the kynurenine pathway
-
-
?
3-hydroxyanthranilate + O2
2-amino-3-carboxymuconate semialdehyde
show the reaction diagram
Pseudomonas fluorescens KU-7
Q83V26
-
-
-
?
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
non-heme enzyme
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
(NH4)2SO4
-
the most effective salt, the optimal concentration is 0.035 M
Fe
-
each monomer contains two iron binding sites. The catalytic iron is buried deep inside the beta-barrel with His51, Glu57, and His95 serving as ligands. The other iron site forms an FeS4 center close to the solvent surface in which the sulfur atoms are provided by Cys125, Cys128, Cys162, and Cys165. The two iron sites are separated by 24 A
Fe2+
-
solvent, acid and heat function to modify the protein configuration so that ferrous ions can be bound to the enzyme to generate the most active form
Fe2+
-
one Fe2+ per active site
Fe2+
-
requirement
Fe2+
-
requirement
Fe2+
-
requirement
Fe2+
-, Q83V26
needed for catalytic activity
Fe2+
-
non-heme ferrous enzyme
Fe2+
-
required, the active site of HAD contains an iron(II) center that is coordinated by the 2-His-1-Glu facial triad
Fe3+
-
does not seem to bind to the enzyme
HCl
-
during purification of enzyme treatment with acid was used
MgCl2
-
-
NaCl
-
-
Ni2+
-
two nickel binding sites per molecule. One of the bound nickel atoms occupies the proposed ferrous-coordinated active site
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-, Q83V26
Fe2+ chelator, 1 mM, complete inhibition
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide methiodide
-, Q83V26
carboxyl-directed reagent, 1 mM, 26% inhibition
2,2'-dipyridyl
-, Q83V26
Fe2+ chelator, 1 mM, complete inhibition
4,6-dibromo-3-hydroxyanthranilic acid
-
NCR-631, characterization of in vivo effects, reversible inhibition with short half-life following systematic administration
4-Bromo-3-hydroxyanthranilic acid
-
competitive
4-Bromo-3-hydroxyanthranilic acid
-
-
4-Chloro-3-hydroxyanthranilate
-
the inactivation results in the consumption of 2 equivalents of oxygen and the production of superoxide. The inhibitor stimulates the oxidation of the active site Fe(II) to the catalytically inactive Fe(III) oxidation state. The inactivated enzyme can be reactivated by treatment with DTT and FeI(II). The nhibitor does not form an adduct with the enzyme. Four conserved cysteines are oxidized to two disulfides (Cys125-Cys128 and Cys162-Cys165) during the inactivation reaction. These results are consistent with a mechanism in which the enzyme, complexed to the inhibitor and O2, generates superoxide which subsequently dissociates, leaving the inhibitor and the oxidized iron center at the active site
4-Chloro-3-hydroxyanthranilate
-
-
4-Chloro-3-hydroxyanthranilate
-
it is possible that inhibition of 3-HAD may improve neurologic status through an increased production of kynurenic acid, a non-specific inhibitor of excitatory amino acid receptors and an inhibitor of quinolinic acid neurotoxicity
4-Chloro-3-hydroxyanthranilic acid
-
competitive
4-Fluoro-3-hydroxyanthranilic acid
-
competitive
6-chloro-3-hydroxyanthranilic acid
-
5-20 mM, loss of enzymatic activity as a function of the inhibitor concentration
acetaminophen
-
-
Acetylsalicylic acid
-
-
anthranilic acid
-
competitive inhibition
Cd2+
-, Q83V26
0.1 mM, more than 99% inhibition
Co2+
-, Q83V26
1 mM, 92% inhibition
Cu2+
-, Q83V26
0.1 mM, complete inhibition
diethyl dicarbonate
-, Q83V26
modifies histidine residues of catechol dioxygenases, 1 mM, 70% inhibition
Dithionitrobenzoic acid
-, Q83V26
cysteine-directed reagent, 1 mM, complete inhibition
EDTA
-, Q83V26
1 mM, 99% inhibition
Fe3+
-, Q83V26
0.1 mM, 82% inhibition
-
Ni2+
-, Q83V26
1 mM, 82% inhibition
o-methoxybenzoylalanine
-
in whole-liver homogenates, but in purified enzyme preparations only in the presence of mitochondria
p-chloromercuribenzoate
-
-
p-Chloromercuriphenyl sulfonic acid
-
-
Quinolinic acid
-
competitive inhibition
Zn2+
-, Q83V26
0.1 mM, 99% inhibition
iodoacetate
-, Q83V26
cysteine-directed reagent, 1 mM, 42% inhibition
additional information
-
not at a concentration of 0.5 mM: L-tryptophan, quinolinic acid, kynurenic acid, nicotinic acid mononucleotide, picolinic acid, phthalic acid, glutaric acid, L-aspartic acid, L-glutamic acid, N-methyl-D-aspartic acid, kainic acid
-
additional information
-
not: Mn2+, Ni2+, Cu2+, Re3+, Os3+, Pb2+
-
additional information
-
both geometrical end electronic structural feature of 4,5- and 4,6-disubstituted and 4,5,6-trisubstituted 3-hydroxyanthranilic derivates play an important role in the inhibitory potency
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Triton X-100
-
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
36
57
3-hydroxy-4-methylanthranilic acid
-
-
0.0102
-
3-Hydroxyanthranilate
-
pH 7.2, mutant enzyme E110A
0.0224
-
3-Hydroxyanthranilate
-
pH 7.2, wild-type enzyme
0.147
-
3-Hydroxyanthranilate
-
pH 7.2, mutant enzyme R47A
0.872
-
3-Hydroxyanthranilate
-
pH 7.2, mutant enzyme R99A
16
19
3-Hydroxyanthranilate
-
-
0.002
-
3-hydroxyanthranilic acid
-
-
0.003
0.0036
3-hydroxyanthranilic acid
-
-
0.0159
-
3-hydroxyanthranilic acid
P47096, -
deleted complemented strain
0.0192
-
3-hydroxyanthranilic acid
P47096, -
wild-type strain
0.021
-
3-hydroxyanthranilic acid
-
-
0.105
-
3-hydroxyanthranilic acid
-
-
0.011
-
4-ethyl-3-hydroxyanthranilic acid
-
-
0.037
-
4-methyl-3-hydroxyanthranilic acid
-
-
0.615
-
O2
-
-
0.01
-
4-Propyl-3-hydroxyanthranilic acid
-
-
additional information
-
additional information
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0046
-
3-Hydroxyanthranilate
-
pH 7.2, mutant enzyme R99A
0.012
-
3-Hydroxyanthranilate
-
pH 7.2, mutant enzyme E110A
0.022
-
3-Hydroxyanthranilate
-
pH 7.2, mutant enzyme R47A
25
-
3-Hydroxyanthranilate
-
pH 7.2, wild-type enzyme
additional information
-
additional information
-
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00004
-
4-Bromo-3-hydroxyanthranilic acid
-
competitive
0.00003
-
4-Chloro-3-hydroxyanthranilic acid
-
competitive
0.00019
-
4-Fluoro-3-hydroxyanthranilic acid
-
competitive
12
-
6-chloro-3-hydroxyanthranilic acid
-
-
2
-
anthranilic acid
-
for the pI 4.98 enzyme
4.9
-
anthranilic acid
-
for the pI 4.98 enzyme
6.5
-
anthranilic acid
-
for the pI 5.6 enzyme
1.8
-
Quinolinic acid
-
for the pI 5.6 enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0003
-
-
brain
0.0075
-
-
liver
6.8
-
-
-
7.8
-
-
-
7.9
-
-, Q83V26
3-hydroxyanthranilate as substrate, pH 6.5, 30C
115.7
-
-
last purification step
140
-
-
-
additional information
-
-
reaches a maximum after 10 min at acid pH
additional information
-
-
-
additional information
-
-
comparison of values of wild-type and deleted complemented strains
additional information
-
P47096, -
comparison of values of wild-type and deleted complemented strains
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
-
-
determination on heat-reactivated extracts after inactivation
6.9
7.4
-
-
7.4
7.6
-
-
additional information
-
-
pI: 5.2
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
7
-
at pH 4.5: about 70% of activity maximum, at pH 7.0: about 50% of activity maximum
5
7.5
-
determination on heat-reactivated extracts after inactivation
additional information
-
-
at pH 3.4: the enzyme is in a form which can bind substrate but is enzymatically inactive, at pH 6.5: active form of enzyme
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
assay at
37
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0
50
-
enzyme activity increased almost linearly with temperature, beyond a sharp drop
55
-
-
activation by heating at 55C for 5 min
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.98
-
-
chromatofocusing in PBE 94 gel exchanger and polybuffer 74, pH 4-7.4
5.6
-
-
chromatofocusing in PBE 94 gel exchanger and polybuffer 74, pH 4-7.4
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
regional distribution
Manually annotated by BRENDA team
-
relatively small amounts
Manually annotated by BRENDA team
Rattus norvegicus Wistar
-
-
-
Manually annotated by BRENDA team
-
activity is low in newborn, significantly incrEases and remains elevated through the following ages
Manually annotated by BRENDA team
-
specific activity progressively and significantly increases from newborns to 12 months of age
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
21240
-
-, Q83V26
predicted from cDNA sequence
23800
-
-, Q83V26
SDS-PAGE
32520
-
-
PAGE in nondenaturing conditions, pI 4.98 enzyme
32570
-
-
PAGE in nondenaturing conditions, pI 5.6 enzyme
32590
-
-
ion spray MS, recombinant enzyme
32600
-
-
immunoblot analysis, recombinant enzyme
32630
-
-
ion spray MS
33000
34000
-
gel filtration
33000
-
-
-
33000
-
-
SDS-PAGE, for the two active enzyme solutions obtained from hydroxyapatite column
33000
-
Q0VCA8
SDS-PAGE, nondenaturating PAGE resolves two components, purified by FPLC on protein PakGlass DEAE-4 PW column, one component is N-terminally truncated annexin IV
34000
-
-
gel filtration, readily aggregates to form inactive higher molecular weight oligomers
35000
40000
-
gel filtration, SDS-PAGE
36000
-
-
SDS-PAGE
37000
38000
-
gel filtration, SDS-PAGE, sucrose density gradient centrifugation
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
monomer
-
1 * 37000-38000, SDS-PAGE
monomer
-
1 * 35000-40000, SDS-PAGE
monomer
-
1* 32589 or 32627, ion spray MS
monomer
-
1 * 33000
monomer
-
1 * 33000-34000, gel filtration
monomer
Q0VCA8
1 * 33000, SDS-PAGE, 286 amino acids, 2 domains that represent the dimers of the prokaryote enzyme structure which is also conserved in simple eukaryotes
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
first vapor diffusion with sitting drop method starting from protein concentration of 10 mg/ml in 32% (NH4)2SO4, 0.1 M sodium acetate, 10 mM 2-mercaptoethanol, pH 5, subsequently, crystals can be obtained by seeding starting from fragments of first crystallization experiments using 40% (NH4)2SO4, Tris-HCl, 40 mM MgCl2, 3% MPD, pH 8 as precipitant
Q0VCA8
hanging drop method
-
hanging drop-vapor diffusion method, 2.4 A resolution
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
-
-
most stable, above extremly unstable
10
-
-
4C, half-life: 3 days
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0
4
-
stable for a month, in Tris-maleate buffer, pH 6.5
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
stability of enzyme in crude extract
-
thawing and refreezing: crude enzyme extract at 20C, relatively stable in presence of Fe2+, about 30% loss of activity after 2 thawings
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
drastic change in the Km of the enzyme in the presence of dimethylglutarate buffer
additional information
-
enzyme is not stable, when stored frozen in 20 mM Tris-maleate buffer, 1 mM dithiothreitol and 1 mM FeSO4 at pH 6.5
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-15C, in 0.01 M Tris buffer, pH 7.0, 4 days, no loss of activity
-
-90C, frozen in dry ice-ethanol bath, partially purified enzyme is stable for at least 1 month, purified enzyme is unstable
-
0C, 0.01 M collidine chloride, 0.01 M potassium chloride, pH 6.5, about 15% loss of activity after 1 month, purified enzyme
-
4C, overnight, about 75% loss of activity
-
0C, 66.7 mM Tris-HCl buffer, pH 8.0, 90 min, approximately 25% loss of activity
-
25C, 66.7 mM Tris-HCl buffer, pH 8.0, 90 min, approximately 70% loss of activity
-
4C, overnight, 75% loss of activity
-
-80C, as homogenate stable for 2 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
bovine kidney homogenized in 0.1 M potassium dihydrogen phosphate buffer with 20% glycerol and 3 mM 2-mercaptoethanol, pH 7.4, centrifuged, protein fraction of supernatant that precipitates from 34-62% saturated (NH4)2SO4 is resuspended in 5 mM potassium dihydrogen phosphate buffer with 20% glycerol and 3 mM 2-mercaptoethanol, pH 7.4, dialyzed against same buffer containing protease inhibitor PMSF (0.1 mM), applied to DEAE Sephadex A-50 column, fractions with enzyme activity pooled and concentrated by precipitation with 80%-saturated (NH4)2SO4, dialyzed against 10 mM Tris-HCl with 20% glycerol and 3 mM 2-mercaptoethanol, pH 7.4, applied to Blue-Sepharose CL-4B column, concentration of active fractions by ultrafiltration through YM-10 membrane
Q0VCA8
succesive size exclusion and affinity columns
-
pure liver enzyme, partielly purified brain enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
expression in Escherichia coli
-, Q83V26
expression in human embryonic kidney HEK-293
-
Y3HAO protein is subcloned into the pET-28a expression vector from extracted Saccharomyces cerevisiae genomic DNA, and the Y3HAO protein is highly expressed as a soluble protein in Escherichia coli strain BL21(DE3) with a six-residueHis tag attached to its N-terminus
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E110A
-
kcat/Km is 954fold lower than wild-type enzyme
R47A
-
kcat/Km is 7440fold lower than wild-type enzyme. Mutant enzyme shows substrate inhibition
R99A
-
kcat/Km is 22320fold lower than wild-type enzyme
H52A
-, Q83V26
inactive enzyme
H93A
-, Q83V26
24.8% activity of the native enzyme
H96A
-, Q83V26
inactive enzyme
H52A
Pseudomonas fluorescens KU-7
-
inactive enzyme
-
H93A
Pseudomonas fluorescens KU-7
-
24.8% activity of the native enzyme
-
H96A
Pseudomonas fluorescens KU-7
-
inactive enzyme
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
plays a role in disorders, associated with altered tissue levels of quinolinic acid