Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli strain DH10B | Klebsiella pneumoniae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
methylated duplex DNA | partial inhibition at high concentrations, mixed inhibition against un- or hemimethylated DNA, noncompetitive against S-adenosyl-L-methionine | Klebsiella pneumoniae | |
S-adenosyl-L-homocysteine | competitive inhibition against S-adenosyl-L-methionine, noncompetitive against DNA | Klebsiella pneumoniae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics, ordered bi bi kinetic mechanism, cooperative binding of 2 molecules of enzyme per DNA is required for activity | Klebsiella pneumoniae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
2 * 45000, recombinant enzyme, SDS-PAGE | Klebsiella pneumoniae |
90000 | - |
recombinant enzyme, gel filtration | Klebsiella pneumoniae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + DNA adenine | Klebsiella pneumoniae | enzyme is a member of a restriction-modification system, R-M system, plasmid DNA, and hemi- or unmethylated duplex DNA, preference for hemimethylated DNA | S-adenosyl-L-homocysteine + DNA 6-methylaminopurine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Klebsiella pneumoniae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli strain DH10B to homogeneity by ultracentrifugation, treatment by 1% v/v polyethyleneimine with 250 mM KCl, ammonium sulfate fractionation, several steps of dialysis, phosphocellulose resin chromatography, and affinity chromatography | Klebsiella pneumoniae |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
S-adenosyl-L-methionine + adenine in DNA = S-adenosyl-L-homocysteine + N6-methyladenine in DNA | ordered bi bi mechanism with S-adenosyl-L-homocysteine leaving at last, cooperative binding of 2 molecules of enzyme per DNA is required for activity | Klebsiella pneumoniae |
Storage Stability | Organism |
---|---|
-20°C, purified recombinant enzyme, 10 mM Tris-HCl, pH 7.4, 0.1 mM EDTA, 100 mM KCl, 7 mM 2-mercaptoethanol, and 10% glycerol, stable for at least 3 months | Klebsiella pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + adenine in DNA | - |
Klebsiella pneumoniae | S-adenosyl-L-homocysteine + N6-methyladenine in DNA | - |
r | |
S-adenosyl-L-methionine + DNA adenine | enzyme is a member of a restriction-modification system, R-M system, plasmid DNA, and hemi- or unmethylated duplex DNA, preference for hemimethylated DNA | Klebsiella pneumoniae | S-adenosyl-L-homocysteine + DNA 6-methylaminopurine | - |
? | |
S-adenosyl-L-methionine + DNA adenine | enzyme recognizes the sequence 5'-GGTACC-3', methylation at position N6, enzyme utilizes plasmid DNA, and hemi- or unmethylated duplex DNA, preference for hemimethylated DNA | Klebsiella pneumoniae | S-adenosyl-L-homocysteine + DNA 6-methylaminopurine | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 45000, recombinant enzyme, SDS-PAGE | Klebsiella pneumoniae |
Synonyms | Comment | Organism |
---|---|---|
KpnI DNA methyltransferase | - |
Klebsiella pneumoniae |
KpnI DNA-(N6-adenine)-methyltransferase | - |
Klebsiella pneumoniae |
KpnI MTase | - |
Klebsiella pneumoniae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Klebsiella pneumoniae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00033 | - |
adenine in DNA | substrate plasmid DNA, recombinant enzyme, pH 8.0, 37°C | Klebsiella pneumoniae | |
0.0011 | - |
adenine in DNA | substrate unmethylated duplex DNA, recombinant enzyme, pH 8.0, 37°C | Klebsiella pneumoniae | |
0.0056 | - |
adenine in DNA | substrate hemimethylated duplex DNA, recombinant enzyme, pH 8.0, 37°C | Klebsiella pneumoniae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Klebsiella pneumoniae |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics | Klebsiella pneumoniae | |
0.00062 | - |
S-adenosyl-L-homocysteine | versus S-adenosyl-L-methionine, pH 8.0, 37°C | Klebsiella pneumoniae |