RESUMEN

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.

Asunto(s)

Alquinos/química , Azidas/química , ADN/análisis , Colorantes Fluorescentes/análisis , S-Adenosilmetionina/análogos & derivados , Alquilación , Alquinos/metabolismo , Azidas/metabolismo , Química Clic , ADN/metabolismo , Colorantes Fluorescentes/metabolismo , Plásmidos/análisis , Plásmidos/metabolismo , S-Adenosilmetionina/metabolismo , Metiltransferasa de ADN de Sitio Específico (Adenina Especifica)/metabolismo