Information on EC 1.14.99.29 - deoxyhypusine monooxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.99.29
-
RECOMMENDED NAME
GeneOntology No.
deoxyhypusine monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
[eIF5A]-deoxyhypusine + AH2 + O2 = [eIF5A]-hypusine + A + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydroxylation
oxidation
redox reaction
-
-
-
-
reduction
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
hypusine biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
deoxyhypusine,hydrogen-donor:oxygen oxidoreductase (2-hydroxylating)
The enzyme catalyses the final step in the formation of the amino acid hypusine in the eukaryotic initiation factor 5A.
CAS REGISTRY NUMBER
COMMENTARY hide
101920-83-6
-
102576-87-4
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of human translation initiation factor 5A + electron donor + O2
6-N-(4-aminobutyl)-L-lysine of human translation initiation factor 5A + oxidized electron donor + H2O
show the reaction diagram
6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of yeast translation initiation factor 5A + electron donor + O2
6-N-(4-aminobutyl)-L-lysine of yeast translation initiation factor 5A + oxidized electron donor + H2O
show the reaction diagram
eIF5A-N6-(4-aminobutyl)-L-lysine + AH2 + O2
eIF5A-N6-(4-amino-2-hydroxybutyl)-L-lysine + A + H2O
show the reaction diagram
eIF5A-Nepsilon-(4-aminobutyl)lysine + AH2 + O2
eIF5A-Nepsilon-(4-amino-2-hydroxybutyl)lysine + A + H2O
show the reaction diagram
eukaryotic translation initiation factor eIF-5A deoxyhypusine + O2 + AH2
eukaryotic translation initiation factor eIF-5A hypusine + H2O + A
show the reaction diagram
N'-(4-aminobutyl)lysine + electron donor + O2
N'-(4-amino-2-hydroxybutyl)lysine + oxidized electron donor + H2O
show the reaction diagram
[eIF5A]-deoxyhypusine + AH2 + O2
[eIF5A]-hypusine + A + H2O
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
eIF5A-N6-(4-aminobutyl)-L-lysine + AH2 + O2
eIF5A-N6-(4-amino-2-hydroxybutyl)-L-lysine + A + H2O
show the reaction diagram
eIF5A-Nepsilon-(4-aminobutyl)lysine + AH2 + O2
eIF5A-Nepsilon-(4-amino-2-hydroxybutyl)lysine + A + H2O
show the reaction diagram
eukaryotic translation initiation factor eIF-5A deoxyhypusine + O2 + AH2
eukaryotic translation initiation factor eIF-5A hypusine + H2O + A
show the reaction diagram
[eIF5A]-deoxyhypusine + AH2 + O2
[eIF5A]-hypusine + A + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
no activity with NADPH, ascorbic acid, mercaptoethylamine, mercaptoacetic acid
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe3+
-
1.5-fold activation at 0.001 mM
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-Diaminodecane
-
94% of initial activity at 2 mM
1,10-phenanthroline
-
complete inhibition at 0.01 mM
1,3-Diaminopropane
-
86% of initial activity at 0.5 mM
1,6-diaminohexane
-
97% of initial activity at 2 mM
1,7-Diaminoheptane
-
91% of initial activity at 2 mM
1,8-diaminooctane
-
93% of initial activity at 2 mM
2,2'-dipyridyl
2,3-Dihydroxybenzoic acid
-
slight inhibition at 2 mM
2-(2-hydroxy-5-methylphenyl)-1,3-thiazole-4-carboxylic acid
-
inhibition in vitro and in cells
2-(2-hydroxy-5-methylphenyl)-4,5-dihydro-1,3-thiazole-4-carboxylic acid
3,4-dihydroxybenzoic acid
-
above 50% inhibition at 2 mM
4,6-diphenyl-1-hydroxy-pyridine-2-one
-
IC50 0.0007 mM
alkyl 4-oxo-piperidine 3 carboxylates
-
structurally related to dihydropyrimidines, most potent, putative DOHH inhibitors in vitro
CaCl2
-
98% of initial activity at 0.005 mM
cadaverine
-
87% of initial activity at 0.5 mM
caldine
-
47% of initial activity at 0.5 mM
ciclopirox
Co(C2H3O2)2
CuCl2
-
above 0.01 mM
deferiprone
deferoxamine
-
targets the active metalloenzyme and inhibits DOHH in human vascular endothelial cells
desferrioxamine B
-
IC50 0.016 mM
desferrioxamine mesylate
-
-
ethyl 3,4-dihydroxybenzoate
-
IC50 0.5 mM
FeCl3
-
65% of initial activity at 0.005 mM
FeSO4
-
13% of initial activity at 0.005 mM
Iron chelators
-
-
-
Lys-Thr-Gly-deoxyhypusine-His-Gly-His-Ala-Lys
-
competitive inhibition
methyl 2,3-dihydroxybenzoate
-
IC50 1.6 mM
metipirox
-
IC50 0.0028 mM
mimosine
Mn(C2H3O2)2
-
above 0.001 mM
MnCl2
-
64% of initial activity at 0.005 mM
N-(2-cyanoethyl)butane-1,4-diamine
-
80% of initial activity at 2 mM
N-(3-cyanopropyl)propane-1,3-diamine
-
79% of initial activity at 2 mM
N1-acetyl-L-Orn-L-Pro-Gly
-
above 2 mM
N1-acetyl-N4-(2,3-dihydroxybenzoyl)-L-Orn-L-Pro-Gly
-
IC50 0.2 mM
N1-acetyl-N4-(3,4-dihydroxybenzoyl)-L-Orn-L-Pro-Gly
-
IC50 0.03 mM
Ni(C2H3O2)2
-
above 0.001 mM
NiSO4
-
72% of initial activity at 0.005 mM
Picolinic acid
putrescine
-
85% of initial activity at 0.5 mM
pyridine 2,3-dicarboxylate
Pyridine 2,4-dicarboxylate
Pyridine 2,5-dicarboxylate
Pyridine 3,4-dicarboxylate
Pyridine 3,5-dicarboxylate
spermidine
-
58% of initial activity at 0.5 mM
spermine
-
41% of initial activity at 0.5 mM
thermine
-
35% of initial activity at 0.5 mM
Zn(C2H3O2)2
-
above 0.01 mM
ZnCl2
-
93% of initial activity at 0.005 mM
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
sulfhydryl compound
additional information
-
the enzyme requires addition of sulfhydryl groups for catalytic activity
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000065
6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of human translation initiation factor 5A
-
pH 7.5, 37C
-
0.000376
6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of yeast translation initiation factor 5A
-
pH 7.5, 37C
-
0.000052
deoxyhypusine
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.44
Lys-Thr-Gly-deoxyhypusine-His-Gly-His-Ala-Lys
-
competitive inhibition
0.032
N1-acetyl-N4-(3,4-dihydroxybenzoyl)-L-Orn-L-Pro-Gly
-
competitive inhibition
0.25
spermine
-
competitive inhibition
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.026 - 0.029
2,2'-dipyridyl
0.0007
4,6-diphenyl-1-hydroxy-pyridine-2-one
Homo sapiens
-
IC50 0.0007 mM
0.0006 - 0.005
ciclopirox
0.117
deferiprone
Homo sapiens
-
IC50 0.117 mM
0.016
desferrioxamine B
Homo sapiens
-
IC50 0.016 mM
0.0003
EDTA
Homo sapiens
-
IC50 0.0003 mM
0.5
ethyl 3,4-dihydroxybenzoate
Rattus norvegicus
-
IC50 0.5 mM
1.6
methyl 2,3-dihydroxybenzoate
Rattus norvegicus
-
IC50 1.6 mM
0.0028
metipirox
Homo sapiens
-
IC50 0.0028 mM
0.0033 - 0.191
mimosine
0.2
N1-acetyl-N4-(2,3-dihydroxybenzoyl)-L-Orn-L-Pro-Gly
Rattus norvegicus
-
IC50 0.2 mM
0.03
N1-acetyl-N4-(3,4-dihydroxybenzoyl)-L-Orn-L-Pro-Gly
Rattus norvegicus
-
IC50 0.03 mM
0.0000125
zileuton
Plasmodium vivax
A5JZ19
recombinant His-tagged enzyme, pH 7.5, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.5
-
in 20 mM Tris buffer
7.4
activity assay
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
activity assay at room temperature
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.9
about, sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
larval garland cell
Manually annotated by BRENDA team
-
larval imaginal disc cell
Manually annotated by BRENDA team
additional information
-
distribution in mammals
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000
-
x * 32000, calculated
33000
x * 33000, about, sequence calculation
36000
-
x * 36000, calculated
39000
-
determined by SDS-PAGE and Western Blot analysis
39010
x * 39010, recombinant His-tagged enzyme, SDS-PAGE
46450
theoretical
additional information
-
The apoenzyme has a larger hydrodynamic size than the holoenzyme
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
20 mM Tris buffer stabilizes
-
freezing/thawing, stable for numerous times
-
sulfhydryl compound necessary
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, stable for at least 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme is partially purified from testis
-
native plasmodial DOHH by anion exchange chromatography and nickel affinity chromatography
-
performed by nickel-chelate affinity chromatography under native conditions
recombinant GST-tagged DOHH by glutathione affinity chromatography
-
recombinant GST-tagged DOHH by glutathione affinity chromatography. Recombinant His-tagged hypusine pathway enzymes from Escherichia coli by anion exchange and nickel affinity chromatography
-
recombinant GST-tagged Lia1 from Escherichia coli strain BL21(DE3) by two steps of glutathione affinity chromatography and gel filtration. Separation of iron-free and iron-bound forms by gel filtration and native electrophoresis
-
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography, the tag is cleaved off by thrombin
-
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain JM109 by glutathione affinity chromatography, the GST-tag is cleaved off
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affiniity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of a polycistronic vector encoding eIF5A, DHS and DOHH, expression in Escherichia coli strain BL21(DE3), overview. Effects of time and temperature on the expression of recombinant proteins and hypusine modification, DOHH effectively hydroxylates all the eIF5A(Dhp), a deoxyhypusine-containing intermediate, formed
-
DNA and amino acid sequence determination and analysis, different clones, sequence comparisons, expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain JM109
expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
for expression in Escherichia coli cells
-
gene dohh, cloned from a GST-ORF library in a biochemical genomics approach. The dohh gene complements a yeast deficient mutant YJR070C, expression as GST-tagged protein
-
gene dohh, quantitative expression analysis
gene DOHH, quantitative real-time PCR expression analysis
-
gene dohh, sequence comparison, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene LIA1, expression in Escherichia coli strain BL21(DE3) as GST-tagged protein
-
into the pST-Acceptor vector for sequencing, into the vectors pET-15b and pET-28a for expression in Escherichia coli BL21DE3 cells
single copy gene dohh
single copy gene dohh, expression as GST-tagged protein, expression of the hypusine pathway genes in Escherichia coli as His-tagged proteins
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
transient overexpression of micro-RNA miR-331-3p and/or miR-642-5p in DU-145 prostate cancer cells reduces DOHH mRNA and protein expression and inhibits cell proliferation
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E57G
site-directed mutagenesis, the substitution located at one of the four conserved His-Glu pairs, the potential metal coordination sites, results in severe reduction of deoxyhypusine hydroxylase activity compared to the wild-type enzyme
E208D
-
impairment in binding of substrate translation initiation factor 5A, retains some activity; site-directed mutagenesis, the mutant shows reduced but not fully abolished eIF5A(deoxyhypusine) binding
E208N
-
site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding
E208Q
-
site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding
E57D
-
impairment in binding of substrate translation initiation factor 5A, retains some activity; site-directed mutagenesis, the mutant shows reduced but not fully abolished eIF5A(deoxyhypusine) binding
E57N
-
site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding
E57Q
-
site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding
E93A
-
site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine) binding
G214A
-
severe impairment in binding of substrate translation initiation factor 5A; site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine) but unaltered iron binding
G247A
-
site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate
G63A
-
severe impairment in binding of substrate translation initiation factor 5A; site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine), but unaltered iron binding
M237A
-
site-directed mutagenesis, the mutant shows reduced, but existing eIF5A(deoxyhypusine) and iron binding, but is catalytically inactive
Q215
-
site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate
R183a
-
site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate
R26A
-
site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate
R88A
-
site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate
S202A
-
site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate
E113A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
E116A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
E116D
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
E238A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
E271A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
E80A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
H112A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
H237A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
H270A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
H79A
-
site-directed mutagenesis, the mutant is completely inactive in deoxyhypusine hydroxylation, structure comparison to the wild-type enzyme
E66K
-
mutation E66K in the DOHH homologue gene, Mmd1, causes a temperature-sensitive growth phenotype and altered mitochondrial morphology and distribution
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
activity of metal-free apoenzyme can be restored by addition of Fe2+, but not by Cd2+, Co2+, Cr2+, Cu2+, Mg2+, Mn2+, Ni2+, Zn2+
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
medicine
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