Fluorescent DNA labeling by N-mustard analogues of S-adenosyl-L-methionine

Chembiochem. 2012 Oct 15;13(15):2225-33. doi: 10.1002/cbic.201200438. Epub 2012 Sep 7.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylation
  • Alkynes / chemistry*
  • Alkynes / metabolism
  • Azides / chemistry*
  • Azides / metabolism
  • Click Chemistry
  • DNA / analysis*
  • DNA / metabolism
  • Fluorescent Dyes / analysis*
  • Fluorescent Dyes / metabolism
  • Plasmids / analysis
  • Plasmids / metabolism
  • S-Adenosylmethionine / analogs & derivatives*
  • S-Adenosylmethionine / metabolism
  • Site-Specific DNA-Methyltransferase (Adenine-Specific) / metabolism

Substances

  • Alkynes
  • Azides
  • Fluorescent Dyes
  • S-Adenosylmethionine
  • DNA
  • Site-Specific DNA-Methyltransferase (Adenine-Specific)