Information on EC 1.14.11.16 - peptide-aspartate beta-dioxygenase

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The expected taxonomic range for this enzyme is: Eutheria

EC NUMBER
COMMENTARY hide
1.14.11.16
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RECOMMENDED NAME
GeneOntology No.
peptide-aspartate beta-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
peptide-L-aspartate + 2-oxoglutarate + O2 = peptide-3-hydroxy-L-aspartate + succinate + CO2
show the reaction diagram
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decarboxylation
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hydroxylation
oxidation
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-
-
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redox reaction
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-
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reduction
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-
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SYSTEMATIC NAME
IUBMB Comments
peptide-L-aspartate,2-oxoglutarate:oxygen oxidoreductase (3-hydroxylating)
Requires Fe2+. Some vitamin K-dependent coagulation factors, as well as synthetic peptides based on the structure of the first epidermal growth factor domain of human coagulation factor IX or X, can act as acceptors.
CAS REGISTRY NUMBER
COMMENTARY hide
122544-66-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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inhibition by fetal alcohol spectrum disorder, decrease leads to impairment in neuronal migration
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ankyrin repeat domain of endogenous Notch receptor L-asparagine + 2-oxoglutarate + O2
ankyrin repeat domain of endogenous Notch receptor 3-hydroxy-L-asparagine + succinate + CO2
show the reaction diagram
ankyrin repeat domain protein
?
show the reaction diagram
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a maximum of three ankyrin repeats enables ankyrin repeat domain proteins as a substrate
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-
?
HIF (L-Asn803) + 2-oxoglutarate + O2
HIF (3-hydroxy-L-Asn803) + succinate + CO2
show the reaction diagram
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accepts HIF-1alpha and HIF-2-alpha as substrate, HIF mutant V802A exhibits 4fold lower substrate activity than native protein, no activity with N803A mutant
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-
?
HIF-1alpha peptide Asp788-Leu822 (L-Asn803) + 2-oxoglutarate + O2
HIF-1alpha peptide Asp788-Leu822 (3-hydroxy-L-Asn803) + succinate + CO2
show the reaction diagram
much lower activity with peptides lacking residues 819-822, 807-822, and 815-822
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-
?
peptide L-asparagine + 2-oxoglutarate + O2
peptide 3-hydroxy-L-asparagine + succinate + CO2
show the reaction diagram
peptide L-aspartate + 2-oxoglutarate + O2
peptide 3-hydroxy-L-aspartate + succinate + CO2
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
peptide L-asparagine + 2-oxoglutarate + O2
peptide 3-hydroxy-L-asparagine + succinate + CO2
show the reaction diagram
peptide L-aspartate + 2-oxoglutarate + O2
peptide 3-hydroxy-L-aspartate + succinate + CO2
show the reaction diagram
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl
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at 1 mM 90% inhibition
ankyrin repeat domain
dimethyloxalylglycine
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Insulin-like growth factor-1
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stimulates peptide-aspartate beta-dioxygenase protein expression and directional motility. Ethanol reduces the stimulation without inhibition of the mRNA expression
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iodoacetamide
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at 1 mM less than 5% activity, 2-oxoglutarate and EDTA protects
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ascorbate
required
additional information
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regulated by insulin and insulin-like growth factor
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.005 - 0.125
2-oxoglutarate
0.003 - 0.093
Fe2+
0.019 - 0.075
first epidermal growth factor-like domain
0.1
HIF-1alpha peptide Asp788-Leu822 (L-asparagine803)
pH 7.8, 30C
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0.09
O2
pH 7.8, 30C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0167 - 0.0317
first epidermal growth factor-like domain
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.59
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8
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7.6
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7.9
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about half-maximal activity at pH 6.5 and 7.9
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
assay at room temperature
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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proliferating ducts
Manually annotated by BRENDA team
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highly expressed
Manually annotated by BRENDA team
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hepatocellular carcinoma
Manually annotated by BRENDA team
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of the fetus
Manually annotated by BRENDA team
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highly expressed in all cholangiocarcinomas
Manually annotated by BRENDA team
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highly expressed
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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cerebellar neuronal cell
Manually annotated by BRENDA team
additional information
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the enzyme is overexpressed in a wide variety of human malignancies such as breast, hepatic, biliary, colon, pulmonary, pancreatic, and neural origin
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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membrane-associated
Manually annotated by BRENDA team
additional information
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L-cell extract
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Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28000
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SDS-PAGE
50700
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sedimentation equilibrium
51030
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MALDI-TOF
52000
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SDS-PAGE, protein with lower molecular weight
110000
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Western blot, post-translational modified protein
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
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1 * 50700, sedimentation equilibrium
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
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peptide-aspartate beta-dioxygenase can be phosphorylated by glycogen synthase kinase 3beta, and high levels of glycogen synthase kinase 3beta activity result in decrease in peptide-aspartate beta-dioxygenase protein, whereas inhibition of the kinase and/or global caspases increases peptide-aspartate beta-dioxygenase protein. Phosphorylation is higher in ethanol-treated compared with control cells
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme in complex with peptides of Notch receptor. Significant conformational changes are required in the ankyrin repeat fold of Notch receptor to enable hydroxylation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, 50 mM Tris-HCl pH 7.2, 1 mg/ml bovine serum albumin, at least 8 weeks stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gel filtration, above 95% purity
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His6 affinity chromatography
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Ni2+-NTA garose affinity chromatography
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partial
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recombinant enzyme
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recombinant enzymes
two enzymes, one 52000 Da, the other 56000 Da
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
co-immunization in Mus musculus, thereafter cell fusion of immunized mice splenic lymphocytes with myouse myeloma cells; expression in Escherichia coli DH5alpha
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expressed in HEK293T cells and in Escherichia coli
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expressed in Mus musculus liver cells
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expressed in Sf9 insect cells as His-tag and GST-fusion protein
expressed in wild-type Escherichia coli and in Escherichia coli BL21
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expression in Escherichia coli BL21
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expression in human CNS neuronal and hepatocellulcar carcinoma cells
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in vitro transcription and translation in the presence of canine pancreas microsomes
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transfected into NIH-3T3 cells
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wild-type and mutant enzymes expressed in Escherichia coli
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wild-type and mutants expressed in Escherichia coli BL21
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
about 2fold increase in expression in double transgenic mice with expression of hepatitis Bx and insulin receptor substrate-1 under a liver-specific promoter
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ethanol-treated cultures
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increase after inhibition of GSK-3beta or global caspases
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overexpression in malignant neoplasms
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C637A
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38% activity of wild-type
C644A
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62% activity of wild-type
C656A
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100% activity of wild-type
C681A
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60% activity of wild-type
C696A
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29% activity of wild-type
G659A
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21% activity of wild-type
G669A
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90% activity of wild-type
H667L
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106% activity of wild-type
H671L
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16% activity of wild-type
H675D
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20% activity of wild-type
H675E
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12% activity of wild-type
H686L
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9% activity of wild-type
P678V
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90% activity of wild-type
R682A
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10% activity of wild-type
R684A
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8% activity of wild-type
R684K
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87% activity of wild-type
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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isolation of human single-chain Fv fragments directed against human aspartyl-asparaginyl beta-hydroxylase. Antibodies show significant binding to recombinant enzyme in ELISA, tumor cell lines, and tumor tissues. They target different domains of the enzyme. A goat-anti-human IgG saporin conjugate can be delivered into tumor cells by antibody 6-22 and elicits cytotoxicity toward the tumor cells in vitro
medicine
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