Purification and stabilization of mouse DNA methyltransferase.
Cytosine methylation within DNA has been implicated in genetic imprinting, X-chromosome inactivation, regulation of tissue-specific gene expression, aging, and cancer. Unfortunately, DNA (cytosine-5)-methyltransferases (EC 2.1.1.37) from various mammalian sources have been difficult to isolate and stabilize, precluding investigations of these critical enzymes. We describe a novel FPLC purification of the 190,000 Mr DNA methyltransferase from mouse Friend erythroleukemia cells. The homogeneous 190 kD Mr form of the enzyme is the only polypeptide detected at various stages of cell growth and has not undergone detectable N-terminal proteolysis.
Xu
G
G
Department of Chemistry, University of California, Santa Barbara 93106.
Flynn
J
J
Glickman
J F
JF
Reich
N O
NO
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002455:Cell Division; D002467:Cell Nucleus / Q000201:enzymology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D007700:Kinetics; D007942:Leukemia, Experimental / Q000201:enzymology*; D051379:Mice; D008970:Molecular Weight; D014407:Tumor Cells, Cultured
D004248:DNA (Cytosine-5-)-Methyltransferases