Application | Comment | Organism |
---|---|---|
analysis | the first Gua is recognized by K9, removal of which abrogates the first base-pair recognition, the flipped target Ade binds to the surface of EcoDam in the absence of S-adenosyl-L-methionine, which illustrates a possible intermediate in the base-flipping pathway, the orphaned Thy residue displays structural flexibility by adopting an extrahelical or intrahelical position where it is in contact to N120 | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
His-tagged EcoDam expressed in HMS174(DE3) cells | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
in complex with cognate DNA at 1.89 A resolution, in the presence of S-adenosyl-L-homocysteine | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
L122A | reduces the size of an aliphatic hydrocarbon side chain, is sufficient to convert EcoDam into a bona fide maintenance MTase with pronounced preference for hemimethylated DNA | Escherichia coli |
additional information | EcoDam K9A variant, shows slightly reduced catalytic activity and DNA binding, shows a loss of specificity at the first base-pair, unable to methylate any of the near-cognate sites, base-flipping of substrates carrying a base-pair substitution at the first position of the target site is more efficient than with wild-type | Escherichia coli |
N120A | loses its p-stacking with Gua1, shows small changes in specificity factor S1 | Escherichia coli |
P134A | high catalytic activity, exhibits only a small reduction in the amplitude of the fluorescence change, but no detectable changes in the kinetics of base-flipping, induces base-flipping of the substrate with altered sequence at the third base-pair | Escherichia coli |
P134G | high catalytic activity, exhibits only a small reduction in the amplitude of the fluorescence change, but no detectable changes in the kinetics of base-flipping, induces base-flipping of the substrate with altered sequence at the third base-pair | Escherichia coli |
R124A | overall reduction in catalytic activity but methylates the near-cognate substrates GATT and GATG faster than the canonical GATC, no base-flipping signal | Escherichia coli |
Y138A | loses its interaction with the O6 atom of Gua1, shows small changes in specificity factor S1 | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0AEE8 | - |
- |
Purification (Comment) | Organism |
---|---|
by Ni2+-affinity chromatography and gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + DNA adenine | - |
Escherichia coli | S-adenosyl-L-homocysteine + DNA 6-methyladenine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Dam | - |
Escherichia coli |
Dam DNA-(adenine-N6)-methyltransferase | - |
Escherichia coli |
DNA adenine methyltransferase | - |
Escherichia coli |
EcoDam | - |
Escherichia coli |