Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0034 | - |
single-stranded DNA-3-methylcytosine | at 25°C, pH not specified in the publication | Escherichia coli | |
0.0054 | - |
single-stranded DNA-1-methyladenine | at 25°C, pH not specified in the publication | Escherichia coli | |
0.0062 | - |
double-stranded DNA-1-methyladenine | at 25°C, pH not specified in the publication | Escherichia coli | |
0.0093 | - |
double-stranded DNA-3-methylcytosine | at 25°C, pH not specified in the publication | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | required for activity, AlkB is a member of the non-heme iron (II) 2-oxoglutarate-dependent dioxygenase superfamily | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
DNA-1,N6-ethenoadenine + 2-oxoglutarate + O2 | Escherichia coli | - |
? | - |
? | |
DNA-1-methylguanine + 2-oxoglutarate + O2 | Escherichia coli | - |
DNA-guanine + formaldehyde + succinate + CO2 | - |
? | |
DNA-3,N4-ethenocytosine + 2-oxoglutarate + O2 | Escherichia coli | - |
? | - |
? | |
DNA-3-methylthymine + 2-oxoglutarate + O2 | Escherichia coli | - |
DNA-thymine + formaldehyde + succinate + CO2 | - |
? | |
double-stranded DNA-1-methyladenine + 2-oxoglutarate + O2 | Escherichia coli | 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 | DNA-adenine + formaldehyde + succinate + CO2 | - |
? | |
double-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2 | Escherichia coli | 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 | DNA-cytosine + formaldehyde + succinate + CO2 | - |
? | |
single-stranded DNA-1-methyladenine + 2-oxoglutarate + O2 | Escherichia coli | 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 | DNA-adenine + formaldehyde + succinate + CO2 | - |
? | |
single-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2 | Escherichia coli | 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 | DNA-cytosine + formaldehyde + succinate + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography and SP-Sepharose cation exchange affinity column chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
DNA-1,N6-ethenoadenine + 2-oxoglutarate + O2 | - |
Escherichia coli | ? | - |
? | |
DNA-1-methylguanine + 2-oxoglutarate + O2 | - |
Escherichia coli | DNA-guanine + formaldehyde + succinate + CO2 | - |
? | |
DNA-3,N4-ethenocytosine + 2-oxoglutarate + O2 | - |
Escherichia coli | ? | - |
? | |
DNA-3-methylthymine + 2-oxoglutarate + O2 | - |
Escherichia coli | DNA-thymine + formaldehyde + succinate + CO2 | - |
? | |
double-stranded DNA-1-methyladenine + 2-oxoglutarate + O2 | 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 | Escherichia coli | DNA-adenine + formaldehyde + succinate + CO2 | - |
? | |
double-stranded DNA-1-methyladenine + 2-oxoglutarate + O2 | 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 | Escherichia coli | double-stranded DNA-adenine + formaldehyde + succinate + CO2 | - |
? | |
double-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2 | 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 | Escherichia coli | DNA-cytosine + formaldehyde + succinate + CO2 | - |
? | |
double-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2 | 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 | Escherichia coli | double-stranded DNA-cytosine + formaldehyde + succinate + CO2 | - |
? | |
single-stranded DNA-1-methyladenine + 2-oxoglutarate + O2 | 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 | Escherichia coli | DNA-adenine + formaldehyde + succinate + CO2 | - |
? | |
single-stranded DNA-1-methyladenine + 2-oxoglutarate + O2 | 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 | Escherichia coli | single-stranded DNA-adenine + formaldehyde + succinate + CO2 | - |
? | |
single-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2 | 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 | Escherichia coli | DNA-cytosine + formaldehyde + succinate + CO2 | - |
? | |
single-stranded DNA-3-methylcytosine + 2-oxoglutarate + O2 | 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 | Escherichia coli | single-stranded DNA-cytosine + formaldehyde + succinate + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AlkB | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.037 | - |
single-stranded DNA-3-methylcytosine | at 25°C, pH not specified in the publication | Escherichia coli | |
0.052 | - |
double-stranded DNA-1-methyladenine | at 25°C, pH not specified in the publication | Escherichia coli | |
0.055 | - |
double-stranded DNA-3-methylcytosine | at 25°C, pH not specified in the publication | Escherichia coli | |
0.062 | - |
single-stranded DNA-1-methyladenine | at 25°C, pH not specified in the publication | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
2-oxoglutarate | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
physiological function | the AlkB protein catalyzes the direct reversal of alkylation damage to DNA, primarily 1-methyladenine and 3-methylcytosine lesions created by endogenous or environmental alkylating agents | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.8 | - |
double-stranded DNA-3-methylcytosine | at 25°C, pH not specified in the publication | Escherichia coli | |
8 | - |
double-stranded DNA-1-methyladenine | at 25°C, pH not specified in the publication | Escherichia coli | |
10.8 | - |
single-stranded DNA-3-methylcytosine | at 25°C, pH not specified in the publication | Escherichia coli | |
11.3 | - |
single-stranded DNA-1-methyladenine | at 25°C, pH not specified in the publication | Escherichia coli |