Information on EC 1.14.11.33 - DNA oxidative demethylase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.11.33
-
RECOMMENDED NAME
GeneOntology No.
DNA oxidative demethylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
DNA-base-CH3 + 2-oxoglutarate + O2 = DNA-base + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
demethylation
oxidative demethylation
SYSTEMATIC NAME
IUBMB Comments
methyl DNA-base, 2-oxoglutarate:oxygen oxidoreductase (formaldehyde-forming)
Contains iron; activity is slightly stimulated by ascorbate. Catalyses oxidative demethylation of the DNA base lesions N1-methyladenine, N3-methylcytosine, N1-methylguanine, and N3-methylthymine. It works better on single-stranded DNA (ssDNA) and is capable of repairing damaged bases in RNA.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
-
ABH2 is a member of the ABH family DNA repair dioxygenases
malfunction
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
1-ethyladenine + O2 + 2-oxoglutarate
adenine + CO2 + acetaldehyde + succinate + H+
show the reaction diagram
-
-
-
-
?
1-methyl-adenine in 5'-dAAAA-1MeA-YYAAA + 2-oxoglutarate + O2
5'-dAAAAAYYAAA + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
1-methyl-dAMP + 2-oxoglutarate + O2
dAMP + formaldehyde + succinate + CO2
show the reaction diagram
1-methyl-dATP + 2-oxoglutarate + O2
dATP + formaldehyde + succinate + CO2
show the reaction diagram
1-methyladenine + O2 + 2-oxoglutarate
adenine + CO2 + formaldehyde + succinate + H+
show the reaction diagram
-
-
-
-
?
2'-deoxy-1-methyl-adenosine 3'-phosphate + 2-oxoglutarate + O2
2'-deoxyadenosine 3'-phosphate + formaldehyde + succinate + CO2
show the reaction diagram
3-methylcytosine + O2 + 2-oxoglutarate
cytosine + CO2 + formaldehyde + succinate + H+
show the reaction diagram
-
-
-
-
?
d(Tpm1A) + 2-oxoglutarate + O2
d(TpA) + formaldehyde + succinate + CO2
show the reaction diagram
d(Tpm1ApT) + 2-oxoglutarate + O2
d(TpApT) + formaldehyde + succinate + CO2
show the reaction diagram
DNA-1,6-ethenoadenine + O2
DNA-adenine + glyoxal
show the reaction diagram
DNA-1,N6-ethenoadenine + 2-oxoglutarate + O2
?
show the reaction diagram
DNA-1,N6-ethenoadenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
DNA-1-(2-hydroxypropyl)-adenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-ethanoadenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-ethenoadenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-ethyladenine + 2-oxoglutarate + O2
DNA-adenine + acetaldehyde + succinate + CO2
show the reaction diagram
DNA-1-hydroxyethyladenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-methyladenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
DNA-1-methylcytosine-CH3 + 2-oxoglutarate + O2
DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
DNA-1-methyldeoxyadenine + 2-oxoglutarate + O2
DNA-deoxyadenine + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
DNA-1-methylguanine + 2-oxoglutarate + O2
DNA-guanine + formaldehyde + succinate + CO2
show the reaction diagram
DNA-2-furfurylguanosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-2-methylguanosine + 2-oxoglutarate + O2
DNA-guanosine + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
DNA-3,N4-ethenocytosine + 2-oxoglutarate + O2
?
show the reaction diagram
DNA-3-methylcytosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-3-methylcytosine + 2-oxoglutarate + O2
DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
DNA-3-methylthymine + 2-oxoglutarate + O2
DNA-thymine + formaldehyde + succinate + CO2
show the reaction diagram
DNA-4-methylcytosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-base-CH3 + 2-oxoglutarate + O2
DNA-base + formaldehyde + succinate + CO2
show the reaction diagram
DNA-N1-methyladenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
highest activity
-
-
?
DNA-tetrahydro-2-furfurylguanosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
double-stranded DNA-1-methyladenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
double-stranded DNA-1-methyladenine + 2-oxoglutarate + O2
double-stranded DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
double-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2
DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
double-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2
double-stranded DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
methylated DNA bacteriophage M13mp18 + 2-oxoglutarate + O2
DNA bacteriophage M13mp18 + formaldehyde + succinate + CO2
show the reaction diagram
methylated double-stranded bacteriophage lambda + 2-oxoglutarate + O2
double-stranded bacteriophage lambda + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
methylated luciferase-mRNA + 2-oxoglutarate + O2
luciferase-mRNA + formaldehyde + succinate + CO2
show the reaction diagram
methylated poly(deoxyadenine) + 2-oxoglutarate + O2
poly(deoxyadenine) + formaldehyde + succinate + CO2
show the reaction diagram
methylated poly(deoxycytosine) + 2-oxoglutarate + O2
poly(deoxycytosine) + formaldehyde + succinate + CO2
show the reaction diagram
methylated poly(deoxythymine) + 2-oxoglutarate + O2
poly(deoxythymine) + formaldehyde + succinate + CO2
show the reaction diagram
methylated RNA bacteriophage MS2 + 2-oxoglutarate + O2
RNA bacteriophage MS2 + formaldehyde + succinate + CO2
show the reaction diagram
methylated single-stranded poly(deoxyadenosine) + 2-oxoglutarate + O2
single-stranded poly(deoxyadenosine) + formaldehyde + succinate + CO2
show the reaction diagram
methylated tRNA-Phe + 2-oxoglutarate + O2
tRNA-Phe + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
methylated tRNA-Phe + 2-oxoglutarate + O2
tRNAPhe + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
N1-methyl-ATP + 2-oxoglutarate + O2
ATP + formaldehyde + succinate + CO2
show the reaction diagram
N1-methyladenine + 2-oxoglutarate + O2
adenine + formaldehyde + succinate + CO2
show the reaction diagram
N1-methyladenine in DNA + 2-oxoglutarate + O2
adenine in DNA + formaldehyde + succinate + CO2
show the reaction diagram
-
strong substrate in ss-DNA
-
-
?
N1-methylguanine in DNA + 2-oxoglutarate + O2
guanine in DNA + formaldehyde + succinate + CO2
show the reaction diagram
-
weak substrate in ds-DNA
-
-
?
N3-methylcytosine + 2-oxoglutarate + O2
cytosine + formaldehyde + succinate + CO2
show the reaction diagram
N3-methylcytosine in DNA + 2-oxoglutarate + O2
cytosine in DNA + formaldehyde + succinate + CO2
show the reaction diagram
-
strong substrate in ss-DNA
-
-
?
N3-methylthymine in DNA + 2-oxoglutarate + O2
thymine in DNA + formaldehyde + succinate + CO2
show the reaction diagram
-
weak substrate in ds-DNA
-
-
?
N6-methyladenine + 2-oxoglutarate + O2
adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
via 6-hydroxymethyl adenine derivative
-
-
?
poly(dm1A) + 2-oxoglutarate + O2
poly(dA) + formaldehyde + succinate + CO2
show the reaction diagram
RNA-1-methyladenine + 2-oxoglutarate + O2
RNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
RNA-3-methylcytosine + 2-oxoglutarate + O2
RNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
RNA-base-CH3 + 2-oxoglutarate + O2
RNA-base + formaldehyde + succinate + CO2
show the reaction diagram
single-stranded DNA-1-methyladenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
single-stranded DNA-1-methyladenine + 2-oxoglutarate + O2
single-stranded DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
single-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2
DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
single-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2
single-stranded DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
tRNA-1-methylguanine + 2-oxoglutarate + O2
tRNA-guanine + formaldehyde + succinate + CO2
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
DNA-1,N6-ethenoadenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1,N6-ethenoadenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
DNA-1-(2-hydroxypropyl)-adenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-ethanoadenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-ethenoadenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-hydroxyethyladenine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-1-methyladenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
DNA-1-methylguanine + 2-oxoglutarate + O2
DNA-guanine + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
DNA-2-furfurylguanosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-2-methylguanosine + 2-oxoglutarate + O2
DNA-guanosine + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
DNA-3,N4-ethenocytosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-3-methylcytosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-3-methylcytosine + 2-oxoglutarate + O2
DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
DNA-3-methylthymine + 2-oxoglutarate + O2
DNA-thymine + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
DNA-4-methylcytosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
DNA-base-CH3 + 2-oxoglutarate + O2
DNA-base + formaldehyde + succinate + CO2
show the reaction diagram
DNA-tetrahydro-2-furfurylguanosine + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
double-stranded DNA-1-methyladenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
double-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2
DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
methylated double-stranded bacteriophage lambda + 2-oxoglutarate + O2
double-stranded bacteriophage lambda + formaldehyde + succinate + CO2
show the reaction diagram
-
-
-
-
?
methylated RNA bacteriophage MS2 + 2-oxoglutarate + O2
RNA bacteriophage MS2 + formaldehyde + succinate + CO2
show the reaction diagram
-
isoform ABH3 reactivates methylated single-stranded RNA bacteriophage MS2
-
-
?
N1-methyladenine + 2-oxoglutarate + O2
adenine + formaldehyde + succinate + CO2
show the reaction diagram
N3-methylcytosine + 2-oxoglutarate + O2
cytosine + formaldehyde + succinate + CO2
show the reaction diagram
N6-methyladenine + 2-oxoglutarate + O2
adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
via 6-hydroxymethyl adenine derivative
-
-
?
RNA-base-CH3 + 2-oxoglutarate + O2
RNA-base + formaldehyde + succinate + CO2
show the reaction diagram
single-stranded DNA-1-methyladenine + 2-oxoglutarate + O2
DNA-adenine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
single-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2
DNA-cytosine + formaldehyde + succinate + CO2
show the reaction diagram
-
major substrate, AlkB demethylates DNA-1-methyladenine and DNA-3-methylcytosine with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-oxoglutarate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
-
the double-stranded DNA preference of isoform ABH2 is observed only in the presence of magnesium, the presence of magnesium stimulates the activity of isoform ABH2 on double-stranded DNA, but inhibits its activity on single-stranded DNA (maximal effect is observed in the presence of 10 mM MgCl2)
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
D-2-hydroxyglutarate
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ascorbate
-
2 m ascorbate is used in assay conditions
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0004
1-methyl-adenine in 5'-dAAAA-1MeA-YYAAA
-
pH not specified in the publication, temperature not specified in the publication
-
0.002
1-methyl-dAMP
-
apparent value, at 37°C, pH not specified in the publication
0.0055
1-methyl-dATP
-
apparent value, at 37°C, pH not specified in the publication
0.012
2'-deoxy-1-methyl-adenosine 3'-phosphate
-
apparent value, at 37°C, pH not specified in the publication
0.0044 - 0.05
d(Tpm1A)
0.0028
d(Tpm1ApT)
-
apparent value, at 37°C, pH not specified in the publication
0.002
DNA-N1-methyladenine
-
wild type enzyme, pH and temperature not specified in the publication
-
0.0062
double-stranded DNA-1-methyladenine
-
at 25°C, pH not specified in the publication
-
0.0093
double-stranded DNA-3-methylcytosine
-
at 25°C, pH not specified in the publication
-
0.01
N1-methyl-ATP
-
apparent value, at 37°C, pH not specified in the publication
0.0014
poly(dm1A)
-
apparent value, at 37°C, pH not specified in the publication
-
0.0054
single-stranded DNA-1-methyladenine
-
at 25°C, pH not specified in the publication
-
0.0034
single-stranded DNA-3-methylcytosine
-
at 25°C, pH not specified in the publication
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.04
1-methyl-adenine in 5'-dAAAA-1MeA-YYAAA
-
pH not specified in the publication, temperature not specified in the publication
-
0.07
1-methyl-dAMP
-
at 37°C, pH not specified in the publication
0.033
1-methyl-dATP
-
at 37°C, pH not specified in the publication
0.012
2'-deoxy-1-methyl-adenosine 3'-phosphate
-
at 37°C, pH not specified in the publication
0.06
d(Tpm1A)
-
at 37°C, pH not specified in the publication
0.12
d(Tpm1ApT)
-
at 37°C, pH not specified in the publication
0.1235
DNA-N1-methyladenine
-
wild type enzyme, pH and temperature not specified in the publication
-
0.052
double-stranded DNA-1-methyladenine
-
at 25°C, pH not specified in the publication
-
0.055
double-stranded DNA-3-methylcytosine
-
at 25°C, pH not specified in the publication
-
0.02
N1-methyl-ATP
-
at 37°C, pH not specified in the publication
0.2
poly(dm1A)
-
at 37°C, pH not specified in the publication
-
0.062
single-stranded DNA-1-methyladenine
-
at 25°C, pH not specified in the publication
-
0.037
single-stranded DNA-3-methylcytosine
-
at 25°C, pH not specified in the publication
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.7
1-methyl-ATP
-
at 37°C, pH not specified in the publication
35
1-methyl-dAMP
-
at 37°C, pH not specified in the publication
11.7
1-methyl-dATP
-
at 37°C, pH not specified in the publication
3.3
2'-deoxy-1-methyl-adenosine 3'-phosphate
-
at 37°C, pH not specified in the publication
43
d(Tpm1ApT)
-
at 37°C, pH not specified in the publication
61.8
DNA-N1-methyladenine
-
wild type enzyme, pH and temperature not specified in the publication
-
8
double-stranded DNA-1-methyladenine
-
at 25°C, pH not specified in the publication
-
5.8
double-stranded DNA-3-methylcytosine
-
at 25°C, pH not specified in the publication
-
143
poly(dm1A)
-
at 37°C, pH not specified in the publication
-
11.3
single-stranded DNA-1-methyladenine
-
at 25°C, pH not specified in the publication
-
10.8
single-stranded DNA-3-methylcytosine
-
at 25°C, pH not specified in the publication
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.5
D-2-hydroxyglutarate
Homo sapiens
-
isoform ALKBH2, pH and temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
-
repair of DNA-3-methylthymine by AlkB is optimal at pH 6.0, but inefficient
6.5
-
repair of DNA-3-methylthymine by ABH3 is optimal at pH 6.5, but inefficient
7
-
assay at
7.6 - 8
8
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
high ABH2 and ABH3 mRNA levels
Manually annotated by BRENDA team
-
high ABH3 mRNA levels
Manually annotated by BRENDA team
-
high ABH2 mRNA levels
Manually annotated by BRENDA team
-
high ABH3 mRNA levels
Manually annotated by BRENDA team
-
high ABH3 mRNA levels
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
diffuse presence of YFP-ABH3 construct
Manually annotated by BRENDA team
-
diffuse presence of YFP-ABH2 construct
Manually annotated by BRENDA team
additional information
-
actinomycin D treatment resulted in nuclear localization of ABH2. DNAse I treatment results in a loss of ABH2 nucleolar localization. ABH2 nucleolar localization is dependent on its association with both DNA and RNA
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AlkB in complex with double-stranded DNA containing 1,N6-ethenoadenine, 3-methylthymine, and 3-methylcytosine, using 100 mM NaCl, 25 mM MgCl2, 100 mM cacodylate (pH 6.5) and 20-24% (w/v) PEG 8K or 4K
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
the iron(II)-bound AlkB is reasonably stable at 4°C in the presence of 10 mM 2-mercaptoethanol at concentrations up to 2 mM
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-cellulose column chromatography, S-Sepharose column chromatography, Ni-NTA column chromatography, and Mono-S cation exchange column chromatography
DEAE-cellulose column chromatography, SP Sepharose cation exchange column chromatography, and Mono-S cation exchange column chromatography
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GSTrap column chromatography
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His-tagged AlkB is purified by Ni-NTA agarose column chromatography, untagged Alkb is purified by DEAE-cellulose column chromatography, Sepharose column chromatography, and Mono S column chromatography
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His-tagged enzyme is purified by Ni-NTA agarose column chromatography; His-tagged enzyme is purified by Ni-NTA agarose column chromatography
Ni-NTA agarose column chromatography
Ni-NTA column chromatography
Ni-NTA column chromatography and Mono-Q column chromatography
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Ni-NTA column chromatography and SP-Sepharose cation exchange affinity column chromatography
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Ni-NTA column chromatography and UnoS column chromatography
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Ni-NTA column chromatography and UnoS column chromatography; Ni-NTA column chromatography and UnoS column chromatography
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TALON metal affinity resin column chromatography
Talon metal affinity resincolumn chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AlkB is expressed in Escherichia coli BL21(DE3) cells
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AlkB overexpression in Escherichia coli AlkB-deficient strains HK82 and BW25113. AlkB mutant strain HK82 is transformed with the low-copy-number expression plasmid pJB658 containing the alkB gene following a toluic acid-inducible promoter. The recombinant AlkB expressed is less active than the natural endogenous form
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expressed in Escherichia BL21-CodonPlus(DE3)-RIPL cells
expressed in Escherichia coli B834lambda (DE3) cells
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expressed in Escherichia coli BL21 Star(DE3)pLysS cells
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expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3) cells and in Sf9 insect cells. ABH3 expressed in insect cells contains no posttranslational modifications but is nevertheless found to be 5fold more active than ABH3 purified from Escherichia coli, suggesting ABH3 may not be correctly folded when purified from Escherichia coli
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expressed in Escherichia coli BL21(DE3) cells and in Sf9 insect cells; expressed in Escherichia coli BL21(DE3) cells and in Sf9 insect cells
expressed in Escherichia coli BL21(DE3) cells; expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BL21-CodonPlus(DE3)-RIPL cells
expressed in Escherichia coli CodonPlus RIPL cells
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expression of GFP-tagged enzyme in HeLa cells, localization in nucleoli due to the genuine nucleolar localization signal sequence. Ectopic expression of wild-type ABH2 leads to a 2.5fold increase of pre-rRNA, whereas ectopic expression of D173A mutant only leads to a marginal increase of pre-rRNA
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isoforms ABH2 and ABH3 are expressed in baculovirus-infected Sf9 insect cells
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isoforms ABH2 and ABH3 are expressed in Sf9 insect cells
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D61A
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the mutant is unable to rescue methylated RNA phage MS2 and is unable to convert 2-oxoglutarate to succinate
H59A
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the mutant is unable to rescue methylated RNA phage MS2 and is unable to convert 2-oxoglutarate to succinate
D133C
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the mutant contains no iron and shows indistinguishable chromatographic behavior from that of wild type enzyme
D135A
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the mutant displays about 5-15% of wild type activity towards 1-methyladenine at pH 7.0
D135I
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the mutant shows increased activity with RNA-N6-methyladenine and reduced activity with DNA-N1-methyladenine compared to the wild type enzyme
D135S
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the mutant displays about 5-15% of wild type activity towards 1-methyladenine at pH 7.0
E136L
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the mutant shows increased activity with RNA-N6-methyladenine and reduced activity with DNA-N1-methyladenine compared to the wild type enzyme
H131C
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the mutant contains no iron and shows indistinguishable chromatographic behavior from that of wild type enzyme
H187C
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the mutant contains no iron and shows indistinguishable chromatographic behavior from that of wild type enzyme
C118A
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the mutant shows wild type activity
C118A/C129A
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the mutant shows wild type activity
C129A
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the mutant shows wild type activity
C129A/H134A
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the mutant shows 65% of wild type activity
C304A
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the mutant shows wild type activity
C304A/C371A
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the mutant shows wild type activity
C371A
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the mutant shows wild type activity
C371A/H372A
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the mutant shows wild type activity
D173A
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site-directed mutagenesis, the mutant maintains nucleolar localization, but is not able to enhance transcription from the rDNA luciferase reporter. eÉctopic expression of wild-type ABH2 leads to a 2.5fold increase of pre-rRNA, whereas ectopic expression of D173A mutant only leads to a marginal increase of pre-rRNA
H113A
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the mutant shows wild type activity
H113A/C118A
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the mutant shows 83% of wild type activity
H113A/C118A/C129A/H134A
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the mutant shows 69% of wild type activity
H113A/C129A
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the mutant shows wild type activity
H131C
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the mutant slightly prefers single-stranded DNA substrates over any double-stranded DNA
H134A
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the mutant shows wild type activity
H171C
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the ABH2 mutant shows indistinguishable chromatographic behaviors from those of the wild type proteins and cross-links efficiently with the single-stranded DNA-1 and DNA3-6
H191C
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the ABH3 mutant shows indistinguishable chromatographic behaviors from those of the wild type proteins and cross-links to DNA-1 much more efficiently than the wild type protein after a 20 h incubation
H231C
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the ABH1 mutant shows indistinguishable chromatographic behaviors from those of the wild type proteins, no cross-link between the mutant protein and the single-stranded and double-stranded DNA probes is observed
H303A
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the mutant shows wild type activity
H303A/C304A
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the mutant shows wild type activity
H372A
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the mutant shows wild type activity
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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development of a fluorogenic probe design (MAQ) that is directly responsive to ALKBH3 repair activity. MAQ uses the fluorescence-quenching properties of 1-methyladenine, removal of the alkyl group results in an more than 10fold light-up signal. The probe is specific for ALKBH3 over its related homologue ALKBH2 and can be used to identify and measure the effectiveness of enzyme inhibitors. The probe functions efficiently in cells, allowing imaging and quantitation of ALKBH3 activity by microscopy and flow cytometry
Show AA Sequence (204 entries)
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