title: DNA methyltransferase assays.
abstract: DNA methyltransferases are important enzymes and their inhibition has many potential applications. The investigation of DNA methyltransferases as well as screening for potential inhibitors requires specialized enzyme assays. In this chapter, we describe three DNA methyltransferase assays, each of them based on a different method: (1) An assay using radioactively labeled AdoMet and biotinylated DNA substrates that is ideal for enzymatic characterization of these enzymes. (2) An assay using bisulfite conversion of in vitro methylated DNA that is ideal to determine details of the methylation pattern introduced by DNA-(cytosine C5)-methyltransferases. (3) A novel fluorescence-coupled, restriction-based assay suitable for high-throughput screening of DNA methyltransferase inhibitors.
authors.affiliation: Biochemistry Lab, School of Engineering and Science, Jacobs University Bremen, Bremen, Germany.
authors.forename: Renata Z
authors.initials: RZ
authors.lastname: Jurkowska
authors.forename: Alexandre
authors.initials: A
authors.lastname: Ceccaldi
authors.forename: Yingying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Paola B
authors.initials: PB
authors.lastname: Arimondo
authors.forename: Albert
authors.initials: A
authors.lastname: Jeltsch
mesh_terms: D000818:Animals; D001710:Biotin / Q000378:metabolism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism*; D019175:DNA Methylation / Q000187:drug effects; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D051379:Mice; D015183:Restriction Mapping; D013447:Sulfites / Q000494:pharmacology
chemical_list: D013447:Sulfites; D001710:Biotin; D004248:DNA (Cytosine-5-)-Methyltransferases; C042345:hydrogen sulfite