Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D181A | site-directed mutagenesis, inactive mutant, mutation abolishes base flipping, D181 seems to contact and stabilize the flipped base, i.e. the intermediate state of the base flipping process | Escherichia coli |
S188A | site-directed mutagenesis, exchange in the loop next to the active site, 7-8fold reduction of kcat, mutant shows 92% of wild-type enzyme activity | Escherichia coli |
T190A | site-directed mutagenesis, mutant shows 75% of wild-type enzyme activity | Escherichia coli |
Y184A | site-directed mutagenesis, mutant shows 1.7% of wild-type enzyme activity | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | quantitative stopped-flow kinetics using 2-aminopurine as a probe to detect base flipping, wild-type and mutant enzymes | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + DNA adenine | Escherichia coli | - |
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes to over 95% purity | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
S-adenosyl-L-methionine + adenine in DNA = S-adenosyl-L-homocysteine + N6-methyladenine in DNA | D181 and Y184 are essential for activity | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | substrate specificity with different oligonucleotides, substrate binding specificity, overview | Escherichia coli | ? | - |
? | |
S-adenosyl-L-methionine + DNA adenine | - |
Escherichia coli | S-adenosyl-L-homocysteine + DNA 6-methylaminopurine | - |
? | |
S-adenosyl-L-methionine + DNA adenine | required substrate recognition target sequence is GATC, occuring base flipping in absence of S-adenosyl-L-methionine is a biphasic process and very fast, but binding of the flipped base into the active site pocket requiring S-adenosyl-L-methionine is slow, active site contains the conserved DPPY motif, whose tyrosine184 residue stacks to the flipped target base | Escherichia coli | S-adenosyl-L-homocysteine + DNA 6-methylaminopurine | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | secondary structure analysis | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
Dam DNA-(adenine-N6)-methyltransferase | - |
Escherichia coli |
Dam DNA-(adenine-N6)-MTase | - |
Escherichia coli |
EcoDam | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |