title: Fluorescent DNA labeling by N-mustard analogues of S-adenosyl-L-methionine.
abstract: Azide and alkyne-functionalized N-mustard analogues of S-adenosyl-L-methionine have been synthesized and were demonstrated to undergo efficient methyltransferase-dependent DNA alkylation by M.TaqI and M.HhaI. Subsequent labeling of the DNA with a fluorophore was carried out using copper-catalyzed azide-alkyne cycloaddition chemistry and was visualized by fluorescence scanning. This work demonstrates the utility of functionalized N-mustard analogues as biochemical tools to study biological methylation and offers a facile way to site-selectively label substrates of DNA methyltransferases.
authors.affiliation: Department of Chemistry, Wake Forest University, 1834 Wake Forest Road, Winston-Salem, NC 27106, USA.
authors.forename: Yuhao
authors.initials: Y
authors.lastname: Du
authors.forename: Charles E
authors.initials: CE
authors.lastname: Hendrick
authors.forename: Krysta S
authors.initials: KS
authors.lastname: Frye
authors.forename: Lindsay R
authors.initials: LR
authors.lastname: Comstock
mesh_terms: D000478:Alkylation; D000480:Alkynes / Q000737:chemistry* / Q000378:metabolism; D001386:Azides / Q000737:chemistry* / Q000378:metabolism; D057930:Click Chemistry; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D010957:Plasmids / Q000032:analysis / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000031:analogs & derivatives* / Q000378:metabolism; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000378:metabolism
chemical_list: D000480:Alkynes; D001386:Azides; D005456:Fluorescent Dyes; D012436:S-Adenosylmethionine; D004247:DNA; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)