PMID:7590323
| Citation |
Finta, C, Sulima, U, Venetianer, P and Kiss, A (1995) Purification of the KpnI DNA methyltransferase and photolabeling of the enzyme with S-adenosyl-L-methionine. Gene 164:65-9 |
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| Abstract |
An Escherichia coli strain overproducing the KpnI DNA methyltransferase (M.KpnI) was constructed by cloning the kpnIM gene downstream from the inducible T7 phage luminal diameter 10 promoter. A method involving three chromatographic steps has been developed to purify M.KpnI to homogeneity. The purified enzyme has a pH optimum around 7.3 and is inhibited by salts. M.KpnI can be photolabeled by UV-irradiation of the enzyme in the presence of S-adenosyl-L-[methyl-3H]methionine ([methyl-3H]AdoMet). Photolabeling results from a specific interaction between M.KpnI and AdoMet, as indicated by the dependence of photolabeling on native enzyme conformation and by the inhibitory effect of the AdoMet analogs, sinefungin and S-adenosyl-L-homocysteine (AdoHcy). |
| Links | |
| Keywords |
Adenosine/analogs & derivatives; Adenosine/pharmacology; Cloning, Molecular; Escherichia coli/genetics; Escherichia coli/metabolism; Molecular Sequence Data; Protein Binding/drug effects; Recombinant Proteins/biosynthesis; Recombinant Proteins/isolation & purification; S-Adenosylhomocysteine/pharmacology; S-Adenosylmethionine/analogs & derivatives; S-Adenosylmethionine/metabolism; S-Adenosylmethionine/radiation effects; Site-Specific DNA-Methyltransferase (Adenine-Specific)/genetics; Site-Specific DNA-Methyltransferase (Adenine-Specific)/isolation & purification; Site-Specific DNA-Methyltransferase (Adenine-Specific)/metabolism; Site-Specific DNA-Methyltransferase (Adenine-Specific)/radiation effects; Ultraviolet Rays |
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