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Information on EC 1.14.11.30 - hypoxia-inducible factor-asparagine dioxygenase and Organism(s) Homo sapiens
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1.14.11.30 hypoxia-inducible factor-asparagine dioxygenaseIUBMB Comments
Contains iron, and requires ascorbate. Catalyses hydroxylation of an asparagine in the C-terminal transcriptional activation domain of HIF-alpha, the alpha subunit of the transcriptional regulator HIF (hypoxia-inducible factor), which reduces its interaction with the transcriptional coactivator protein p300. The requirement of oxygen for the hydroxylation reaction enables animals to respond to hypoxia.
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Word Map
- 1.14.11.30
-
prolyl
- hydroxylases
-
transactivation
-
oxygen-dependent
- ankyrin
-
normoxia
-
hippel-lindau
-
oxygen-sensing
-
hif-alpha
-
prolyl-hydroxylase
-
fih-mediated
-
2og-dependent
-
hypoxia-sensitive
- dimethyloxalylglycine
-
2-oxoglutarate-dependent
- medicine
- drug development
The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
Synonyms
hif1an, asparaginyl hydroxylase, factor inhibiting hif-1, factor-inhibiting hif, asparaginyl-hydroxylase, hypoxia-inducible factor 1-alpha inhibitor, hif asparagine hydroxylase, hypoxia-inducible factor asparagine hydroxylase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
FIH
SYSTEMATIC NAME
IUBMB Comments
hypoxia-inducible factor-L-asparagine, 2-oxoglutarate:oxygen oxidoreductase (4-hydroxylating)
Contains iron, and requires ascorbate. Catalyses hydroxylation of an asparagine in the C-terminal transcriptional activation domain of HIF-alpha, the alpha subunit of the transcriptional regulator HIF (hypoxia-inducible factor), which reduces its interaction with the transcriptional coactivator protein p300. The requirement of oxygen for the hydroxylation reaction enables animals to respond to hypoxia.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN + 2-oxoglutarate + O2
DESGLPQLTSYDAEV-(3S)-3-hydroxy-L-asparaginyl-APIQGSRNLLQGEELLRALDQVN + succinate + CO2
-
-
-
?
DESGLPQLTSYDCEVNAPI + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 788-806. 9% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
DESGLPQLTSYDCEVNAPIQGSR + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 788-810. 15% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
DESGLPQLTSYDCEVNAPIQGSRNLLQ + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 788-814. 37% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
DESGLPQLTSYDCEVNAPIQGSRNLLQGEEL + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 788-818. 26% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 788-822
-
-
?
ESYLLPELTRYDCEVNVPVLGSSTLLQGGDLLRAL + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-2alpha peptide 832-857. 7% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
hypoxia-inducible factor-L-asparagine peptide + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine peptide + succinate + CO2
-
39-residue peptide corresponding to HIF-1alpha788-826 mutant C800A
hydroxylation at Asn803
-
?
hypoxia-inducible factor1alpha C-terminal oxygen dependent degradation domain-L-proline + 2-oxoglutarate + O2
hypoxia-inducible factor1alpha C-terminal oxygen dependent degradation domain-trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
-
?
hypoxia-inducible factor1alpha N-terminal oxygen dependent degradation domain-L-proline + 2-oxoglutarate + O2
hypoxia-inducible factor1alpha N-terminal oxygen dependent degradation domain-trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
-
?
hypoxia-inducible factor2alpha C-terminal oxygen dependent degradation domain-L-proline + 2-oxoglutarate + O2
hypoxia-inducible factor2alpha C-terminal oxygen dependent degradation domain-trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
-
?
hypoxia-inducible factor2alpha N-terminal oxygen dependent degradation domain-L-proline + 2-oxoglutarate + O2
hypoxia-inducible factor2alpha N-terminal oxygen dependent degradation domain-trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
-
?
LTRYDCEVNVPVLGSSTLL + O2
?
-
hypoxia-inducible factor-2alpha peptide 839-866. 1% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
LTSYDCEVNAPIQGSRNLL + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 795-813. 4% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
rabankyrin-5 + 2-oxoglutarate + O2
trihydroxy-rabankyrin-5 + succinate + CO2
-
hydroxylation at N316, N485 and N649
-
-
?
additional information
?
-
-
the subtrate contains a C-terminal and a N-terminal oxygen-dependent degradation domain, as well as a C-terminal transactivation domain
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
-
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
activity of the hypoxia-inducible factor (HIF) complex is controlled by oxygen-dependent hydroxylation of prolyl and asparaginyl residues. Hydroxylation of specific prolyl residues by 2-oxoglutarate-dependent oxygenases mediates ubiquitinylation and proteasomal destruction of HIF-alpha. Hydroxylation of an asparagine residue (ASn803) in the C-terminal transactivation domain of HIF-alpha abrogates interaction with p300, preventing transcriptional activation
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
the activity of hypoxia-inducible transcription factor HIF, an alphabeta heterodimer that has an essential role in adaptation to low oxygen availability, is regulated by two oxygen-dependent hydroxylation events. Hydroxylation of specific proline residues by HIF prolyl 4-hydroxylases targets the HIF-alpha subunit for proteasomal destruction, whereas hydroxylation of an asparagine in the C-terminal transactivation domain prevents its interaction with the transcriptional coactivator p300
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
the enzyme requires particularly long peptide substrates, so that omission of only a few residues from the N or C terminus of a 35-residue HIF-1alpha sequence markedly reduces its substrate activity. Hydroxylation of two HIF-2alpha peptides is far less efficient than that of the corresponding HIF-1alpha peptides
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
the oxygen in the alcohol of the hydroxyasparagine residue is directly derived from dioxygen
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
FIH hydroxylates Asn803 of HIF-1alpha
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
FIH hydroxylates at the asparaginyl residue in the FIH transcriptional activation domain C-TAD
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
HIF-1alpha
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
HIF1alpha substrate domain structure, overview. Usage of recombinantly expressed HIFalpha proteins spanning the CODDD region, His6-GB1-TEV-HIF1alpha-(498-603) or His6-GB1-TEV-HIF2alpha-(467-575). The enzyme hydroxylates Asn803 of HIF1alpha and Asn847 of HIF2alpha
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
hydroxylation at Asn803
-
-
?
PSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 775-826. 120% of the activity obtained with the 35-amino-acid HIF-1alpha peptide DES35 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL)
-
-
?
PSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN + 2-oxoglutarate + O2
?
-
hypoxia-inducible factor-1alpha peptide 775-826. Mutation of Asn803 in GST-HIF-1alpha-(775826) to alanine, glutamine or glutamate abolishes activity, while an Asp803 mutant still supports some 2-oxoglutarate turnover, at a maximum of 7% of the analogous Asn-803 substrate
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
-
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
activity of the hypoxia-inducible factor (HIF) complex is controlled by oxygen-dependent hydroxylation of prolyl and asparaginyl residues. Hydroxylation of specific prolyl residues by 2-oxoglutarate-dependent oxygenases mediates ubiquitinylation and proteasomal destruction of HIF-alpha. Hydroxylation of an asparagine residue (ASn803) in the C-terminal transactivation domain of HIF-alpha abrogates interaction with p300, preventing transcriptional activation
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
the activity of hypoxia-inducible transcription factor HIF, an alphabeta heterodimer that has an essential role in adaptation to low oxygen availability, is regulated by two oxygen-dependent hydroxylation events. Hydroxylation of specific proline residues by HIF prolyl 4-hydroxylases targets the HIF-alpha subunit for proteasomal destruction, whereas hydroxylation of an asparagine in the C-terminal transactivation domain prevents its interaction with the transcriptional coactivator p300
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
FIH hydroxylates Asn803 of HIF-1alpha
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
FIH hydroxylates at the asparaginyl residue in the FIH transcriptional activation domain C-TAD
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
-
HIF-1alpha
-
-
?
hypoxia-inducible factor-L-asparagine + 2-oxoglutarate + O2
hypoxia-inducible factor-(3S)-3-hydroxy-L-asparagine + succinate + CO2
hydroxylation at Asn803
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Fe2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1-(5-chloro-6-(trifluoromethoxy)-1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid
-
i.e. JNJ-42041935, 2-oxoglutarate analogue
-
H2O2
-
peroxide rapidly inhibits hydroxlation of diverse FIH substrates and inhibits FIH in a range of cell types. Preferential inhibition of N803-hydroxylation compared with P402/P564 hydroxylation by PHDs, EC 1.14.11.29. Cysteine 800 in HIF-1alpha does not regulate N803 or N847 hydroxylation. FIH activity is not restored by exogenous Fe2+
N-(methoxyoxoacetyl)-glycine methyl ester
-
a pan-hydroxylase inhibitor, in vitro and in vivo inhibition
additional information
-
the Ki-value for 3-hydroxypyridine-2-carbonylglycine and N-((3-hydroxy-6-chloroquinolin-2-yl)carbonyl)glycine are above 0.3 mM
-
additional information
-
construction of peptide inhibitors consist of amino acids identical to those in the HIF1alpha C-terminal oxygen-dependent degradation domain residues 556-575 except for the 564 proline residue: DLDLEALA-L-trans-4-fluoroproline-YIPADDDFQLR, DLDLEALA-L-trans-4-hydroxyproline-YIPADDDFQLR, DLDLEALA-L-piperidine-2-carboxylic acid-YIPADDDFQLR, DLDLEALA-L-3,4-dehydroproline-YIPADDDFQLR, and DLDLEALA-L-4-thioproline-YIPADDDFQLR. All peptide inhibitors show specific inhibition of PHD2, EC 1.14.11.29, and no inhibition of FIH
-
additional information
-
temporal dynamics of hydroxylase inhibition, overview
-
additional information
-
screening of iron chelators pyridines, hydroxypyrones/hydroxypyridinones, and catechols as inhibitors for FIH, analysis of selectivity of the inhibitors for FIH compared to PHD2, EC 1.14.11.29. Ligand binding kinetics and structural analysis, overview. Representative inhibitors bind to the metal center in both FIH as an 2-oxoglutarate mimic
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ferritin heavy chain
i.e. FTH1, directly activates FIH to hydroxylate HIF-1alpha to ensure its normoxic inactivation. In iron-rich conditions, the transcriptional activity of HIF-1alpha is also inhibited due to forceful induction of FTH1
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
steady-state kinetic analysis and substrate selectivity for hypoxia-inducible factor and 2-oxoglutarate
-
0.025
2-oxoglutarate
-
pH 7.8, 37°C, Km-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
0.07
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
mutant enzyme D210E, at pH 7.0 and 37°C
-
0.071
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
mutant enzyme D210G, at pH 7.0 and 37°C
-
0.078
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
wild type enzyme, at pH 7.0 and 37°C
-
0.09
O2
-
pH 7.8, 37°C. The Km of FIH for O2 is about 40% of its atmospheric concentration, Km-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
the catalytic center activities obtained for the enzyme purified by two alternative procedures are 85-135 and 70-120 mol/mol/min, respectively
-
0.027
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
mutant enzyme D210E, at pH 7.0 and 37°C
-
0.383
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
mutant enzyme D210G, at pH 7.0 and 37°C
-
0.55
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
wild type enzyme, at pH 7.0 and 37°C
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.37
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
mutant enzyme D210E, at pH 7.0 and 37°C
-
5.7
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
mutant enzyme D210G, at pH 7.0 and 37°C
-
7
DESGLPQLTSYDAEVNAPIQGSRNLLQGEELLRALDQVN
wild type enzyme, at pH 7.0 and 37°C
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.01
3,4-dihydroxybenzoate
-
pH 7.8, 37°C, Ki-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
0.1
DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL
-
pH 7.8, 37°C, Ki-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
0.16
ESYLLPELTRYDCEVNVPVLGSSTLLQGGDLLRAL
-
pH 7.8, 37°C, Ki-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
0.002
oxalylglycine
-
pH 7.8, 37°C, Ki-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
0.1
PSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN
-
pH 7.8, 37°C, Ki-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
-
0.03
Pyridine-2,4-dicarboxylate
-
pH 7.8, 37°C, Ki-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
0.05
Pyridine-2,5-dicarboxylate
-
pH 7.8, 37°C, Ki-value is determined using soluble extracts of cells expressing enzyme-FLAGHis
additional information
additional information
-
the Ki-value for 3-hydroxypyridine-2-carbonylglycine and N-((3-Hydroxy-6-chloroquinolin-2-yl)carbonyl)glycine are above 0.3 mM
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
the enzyme belongs to the 2-oxoglutarate- and iron-dependent dioxygenase family of enzymes
malfunction
metabolism
physiological function
additional information
malfunction
-
FIH silencing leads to spacial displacement of the expression pattern of HIF target genes that depend on the C-TAD, such as carbonic anhydrase IX, to more oxygenated areas, whereas overexpression of FIH restricts this pattern to more hypoxic areas. Silencing of gene fih severely reduced in vitro cell proliferation and in vivo tumor growth of LS174 colon adenocarcinoma and A375 melanoma cells. Silencing of fih also significantly increases both the total and phosphorylated forms of the tumor suppressor p53, leading to an increase in its direct target, the cell cycle inhibitor p21. p53-deficient or mutant cells are totally insensitive to FIH expression. FIH activity is essential for tumor growth through the suppression of the p53-p21 axis, the major barrier that prevents cancer progression
malfunction
-
silencing FIH under conditions where prolyl hydroxylases, EC 1.14.11.29/30, are inhibited results in increased HIF-1alpha transcriptional activity, but paradoxically decreases HIF-1alpha stability. Residual activity of FIH in hypoxia
malfunction
-
the lower enzyme expression in tumor is associated with incomplete tumor encapsulation, vascular invasion, and microvascular density
metabolism
-
HIF transcriptional activity is controlled by the asparaginyl hydroxylase factor inhibiting HIF-1. Hypoxia-induced HIF signalling, mathematical modelling of the pathway, temporal dynamics of the HIF response to hypoxia, and molecular interaction map for the HIF network, overview. The hypoxia inducible factor is switched on and promotes adaptation to hypoxia by upregulating genes involved in angiogenesis, erythropoiesis and glycolysis
metabolism
-
hypoxia and oxidant stress can interact functionally as distinct regulators of HIF transcriptional output involving the enzyme. Oxidant stress activates hypoxia pathways through the inactivation of the oxygen-sensing hypoxia-inducible factor prolyl and asparaginyl hydroxylases
metabolism
-
optimal HIF-1alpha transcriptional activity requires sequential inhibition of both prolyl- and asparaginyl-hydroxylases
physiological function
-
there are at least two major steps involved in the hypoxic induction of the HIF proteins: (i) inhibition of oxygen-dependent hydroxylation on Pro residues in the oxygen-dependent degradation domain to prevent interaction of HIF with the von Hippel-Lindau tumor suppressor/ubiquitin ligase complex and thus avoid proteasomal destruction, and (ii) inhibition of oxygen-dependent hydroxylation of Asn in the COOH-terminal transactivation domain to promote interaction with the p300/CBP coactivator and induce transcription
physiological function
-
FIH modulates the profile of hypoxia-inducible factor downstream genes and of hypoxia-inducible factor target genes in a physiological oxygen gradient: the in vitro spheroid model, overview. FIH enhances tumorigenesis and controls in vitro p53 expression. Factor-inhibiting hypoxia-inducible factor monitors the expression of a spectrum of genes that are dictated by the cell's partial oxygen pressure. This action is mediated by the C-TAD, one of two transactivation domains of the hypoxia-inducible factor
physiological function
-
HIF transcriptional activity is controlled by the asparaginyl hydroxylase factor inhibiting HIF-1
physiological function
-
key enzyme in activation of the hypoxia-inducible factor (HIF) pathway, a critical step in the transcriptional response to hypoxia, role of FIH in hydroxylase regulation of HIF-1alpha, overview. The enzyme is involved in the HIF-1alpha signalling network, overview. Asparaginyl hydroxylation confers upon HIF-1alpha resistance to proteosomal degradation, but the removal of the asparaginyl hydroxylation step is necessary for HIF-1alpha activity
physiological function
-
the enzyme inhibits the hypoxia-inducible factor transcription activation through asparagine hydroxylation
physiological function
-
the FIH hydroxylase is a cellular peroxide sensor that modulates HIF transcriptional activity. Cysteine 800 in HIF-1alpha does not regulate N803 or N847 hydroxylation
physiological function
ferritin heavy chain FTH1 directly interacts with FIH. FTH1 facilitates the FIH-mediated Asn803 hydroxylation in hypoxia-inducible factor HIF-1alpha and prevents the recruitment of p300 to HIF-1alpha through the Asn803 hydroxylation. FTH1 represses the transcriptional activity of HIF-1alpha in HCT-116 cells under either normoxic or hypoxic conditions and downregulates the expression of the HIF-1 target genes
additional information
-
HIF asparaginyl hydroxylase is strikingly more sensitive to peroxide than the HIF prolyl hydroxylases, EC 1.14.11.29. Inhibition of FIH by peroxide persists in hypoxia
additional information
-
modeling of the dynamic regulation of HIF-1alpha transcriptional activity by the hydroxylase. HIF-1alpha stabilisation and transcriptional activity is dependent on oxygen tension
additional information
-
PHD, EC 1.14.11.29, has a higher affinity for oxygen than FIH
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). HIF1N_HUMAN
349
0
40285
Swiss-Prot
other Location (Reliability: 2)
PDB
SCOP
CATH
UNIPROT
ORGANISM
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
formation of an Fe-Cl cofactor structure in variant D201G. The variant binds Fe with a His2Cl facial triad and activates O2 only in the presence of specific anions. A full His2X triad is an absolute requirement for O2 activation
hanging drop vapor diffusion method, using 0.1 M HEPES, 1.2 M (NH4)2SO4, and 3% PEG 400, pH 7.0
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D201E
the mutant shows greatly impaired activity compared to the wild type enzyme
D201G
D201G
removal of the Asp ligand in the His2Asp facial triad, leads to a highly active enzyme. The variant supports anion dependent peptide hydroxylation, exogenous ligand binding to form a complete His2X facial triad is essential for O2 activation
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant dox-inducible FLAG-tagged FIH from U2-OS cells by immunoaffinity chromatography
-
recombinant His- and MBP-tagged FIH from Escherichia coli strain BL21(DE3)pRR692 by nickel affinity and anion exchange chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of N-terminally His-tagged FIH-1 in Escherichia coli strain BL21(DE3)
-
HIF asparaginyl hydroxylase (FIH), His-FIH, FIH-FLAGHis, FIH-V5His, and GST-FIH polypeptides are expressed in Spodoptera frugiperda Sf9 cells
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
-
specific protocols for the knockdown and inhibition of the HIF prolyl hydroxylases 1-3 and the asparagine hydroxylase factor-inhibiting HIF using RNA interference and hydroxylase inhibitors, respectively
drug development
-
the unique function of FIH makes it a prime target for selective inhibition leading to regulatory control of diseases such as cancer and stroke
medicine
-
the high constitutive activities of the proteins with both Pro and Asn substitutions confirm that the relevant prolyl and asparaginyl hydroxylases are attractive targets for therapeutic regulation of hypoxia-inducible factor-1alpha and hypoxia-inducible factor-2alpha
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Koivunen, P.; Hirsilae, M.; Guenzler, V.; Kivirikko, K.I.; Myllyharju, J.
Catalytic properties of the asparaginyl hydroxylase (FIH) in the oxygen sensing pathway are distinct from those of its prolyl 4-hydroxylases
J. Biol. Chem.
279
9899-9904
2004
Homo sapiens
Hewitson, K.S.; McNeill, L.A.; Riordan, M.V.; Tian, Y.M.; Bullock, A.N.; Welford, R.W.; Elkins, J.M.; Oldham, N.J.; Bhattacharya, S.; Gleadle, J.M.; Ratcliffe, P.J.; Pugh, C.W.; Schofield, C.J.
Hypoxia-inducible factor (HIF) asparagine hydroxylase is identical to factor inhibiting HIF (FIH) and is related to the cupin structural family
J. Biol. Chem.
277
26351-26355
2002
Homo sapiens
Lando, D.; Peet, D.J.; Whelan, D.A.; Gorman, J.J.; Whitelaw, M.L.
Asparagine hydroxylation of the HIF transactivation domain: A hypoxic switch
Science
295
858-861
2002
Homo sapiens
Pappalardi, M.B.; McNulty, D.E.; Martin, J.D.; Fisher, K.E.; Jiang, Y.; Burns, M.C.; Zhao, H.; Ho, T.; Sweitzer, S.; Schwartz, B.; Annan, R.S.; Copeland, R.A.; Tummino, P.J.; Luo, L.
Biochemical characterization of human HIF hydroxylases using HIF protein substrates that contain all three hydroxylation sites
Biochem. J.
436
363-369
2011
Homo sapiens
Kwon, H.S.; Choi, Y.K.; Kim, J.W.; Park, Y.K.; Yang, E.G.; Ahn, D.R.
Inhibition of a prolyl hydroxylase domain (PHD) by substrate analog peptides
Bioorg. Med. Chem. Lett.
21
4325-4328
2011
Homo sapiens
Cavadas, M.A.; Nguyen, L.K.; Cheong, A.
Hypoxia-inducible factor (HIF) network: insights from mathematical models
Cell Commun. Signal.
11
42
2013
Homo sapiens
Masson, N.; Singleton, R.S.; Sekirnik, R.; Trudgian, D.C.; Ambrose, L.J.; Miranda, M.X.; Tian, Y.M.; Kessler, B.M.; Schofield, C.J.; Ratcliffe, P.J.
The FIH hydroxylase is a cellular peroxide sensor that modulates HIF transcriptional activity
EMBO Rep.
13
251-257
2012
Homo sapiens
Nguyen, L.K.; Cavadas, M.A.; Scholz, C.C.; Fitzpatrick, S.F.; Bruning, U.; Cummins, E.P.; Tambuwala, M.M.; Manresa, M.C.; Kholodenko, B.N.; Taylor, C.T.; Cheong, A.
A dynamic model of the hypoxia-inducible factor 1alpha (HIF-1alpha) network
J. Cell Sci.
126
1454-1463
2013
Homo sapiens
Flagg, S.C.; Martin, C.B.; Taabazuing, C.Y.; Holmes, B.E.; Knapp, M.J.
Screening chelating inhibitors of HIF-prolyl hydroxylase domain 2 (PHD2) and factor inhibiting HIF (FIH)
J. Inorg. Biochem.
113
25-30
2012
Homo sapiens
Pelletier, J.; Dayan, F.; Durivault, J.; Ilc, K.; Pecou, E.; Pouyssegur, J.; Mazure, N.M.
The asparaginyl hydroxylase factor-inhibiting HIF is essential for tumor growth through suppression of the p53-p21 axis
Oncogene
31
2989-3001
2012
Homo sapiens
Bishop, T.; Ratcliffe, P.J.
HIF hydroxylase pathways in cardiovascular physiology and medicine
Circ. Res.
117
65-79
2015
Homo sapiens
Chan, M.C.; Ilott, N.E.; Schoedel, J.; Sims, D.; Tumber, A.; Lippl, K.; Mole, D.R.; Pugh, C.W.; Ratcliffe, P.J.; Ponting, C.P.; Schofield, C.J.
Tuning the transcriptional response to hypoxia by inhibiting hypoxia-inducible factor (HIF) prolyl and asparaginyl hydroxylases
J. Biol. Chem.
291
20661-20673
2016
Homo sapiens
Hangasky, J.A.; Taabazuing, C.Y.; Martin, C.B.; Eron, S.J.; Knapp, M.J.
The facial triad in the alpha-ketoglutarate dependent oxygenase FIH A role for sterics in linking substrate binding to O2 activation
J. Inorg. Biochem.
166
26-33
2017
Homo sapiens (Q9NWT6)
Ma, M.; Hua, S.; Li, G.; Wang, S.; Cheng, X.; He, S.; Wu, P.; Chen, X.
Prolyl hydroxylase domain protein 3 and asparaginyl hydroxylase factor inhibiting HIF-1 levels are predictive of tumoral behavior and prognosis in hepatocellular carcinoma
Oncotarget
8
12983-13002
2017
Homo sapiens
Jin, P.; Kang, J.; Lee, M.K.; Park, J.W.
Ferritin heavy chain controls the HIF-driven hypoxic response by activating the asparaginyl hydroxylase FIH
Biochem. Biophys. Res. Commun.
499
475-481
2018
Homo sapiens (Q9NWT6)
Chaplin, V.; Hangasky, J.; Huang, H.; Duan, R.; Maroney, M.; Knapp, M.
Chloride supports O2 activation in the D201G facial triad variant of factor-inhibiting hypoxia inducible factor, an alpha-ketoglutarate dependent oxygenase
Inorg. Chem.
57
12588-12595
2018
Homo sapiens (Q9NWT6)
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