Purification and properties of the P15 specific restriction endonuclease from Escherichia coli.
The specific restriction endonuclease of the Escherichia coli plasmid, P15, has been purified to apparent homogeneity by a procedure that includes DNA-cellulose chromatography as well as a new endonuclease assay. Sedimentation on glycerol gradients showed two peaks of activity with values of 11.3 S and 15.7 S. The highly purified enzyme requires ATP and Mg2+ for activity and is stimulated by S-adenosylmethionine. A methylase activity is observed in the course of the endonucleolytic reaction which protects some of the DNA sites from cleavage.
D000251:Adenosine Triphosphatases / Q000302:isolation & purification / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000302:isolation & purification / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000302:isolation & purification / Q000378:metabolism; D004720:Endonucleases / Q000302:isolation & purification; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D012436:S-Adenosylmethionine / Q000494:pharmacology
D012436:S-Adenosylmethionine; D000255:Adenosine Triphosphate; D004248:DNA (Cytosine-5-)-Methyltransferases; D004720:Endonucleases; D004262:DNA Restriction Enzymes; D000251:Adenosine Triphosphatases; D008274:Magnesium