Novel method to detect DNA methylation using gold nanoparticles coupled with enzyme-linkage reactions

Anal Chem. 2010 Jan 1;82(1):229-33. doi: 10.1021/ac902198v.

Abstract

DNA methylation, catalyzed by methylases, plays a critical role in many biological processes, and methylases have been regarded as promising targets for antimicrobial drugs. In this paper, we propose a simple and sensitive colorimetric assay method to detect the activity of methylases so as to monitor DNA methylation using DNA-modified gold nanoparticles (AuNPs) coupled with enzyme-linkage reactions. The duplex DNA molecules modified on the surface of AuNPs are first methylated by DNA adenine methylation (Dam) methyltransferase (MTase) and then cut by methylation-sensitive restriction endonuclease Dpn I. Removal of duplex from the AuNP surfaces by the methylation/cleavage process will destabilize the nanoparticles, resulting in aggregation of AuNPs and a red-to-blue color change. Consequently, the enzyme activity of Dam MTase can be assayed and DNA methylation can be detected. Furthermore, this study may provide a sensitive platform to screen inhibitors for Dam MTase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Colorimetry / methods*
  • DNA / chemistry*
  • DNA Methylation*
  • DNA Modification Methylases
  • Deoxyribonuclease I
  • Gold / chemistry*
  • Metal Nanoparticles / chemistry*
  • Sensitivity and Specificity

Substances

  • Gold
  • DNA
  • DNA Modification Methylases
  • Deoxyribonuclease I