A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters

Anal Bioanal Chem. 2018 Aug;410(20):4943-4952. doi: 10.1007/s00216-018-1143-2. Epub 2018 Jun 22.

Abstract

DNA methylation catalyzed by DNA methyl transferase (MTase) is a significant epigenetic process for modulating gene expression. Abnormal levels of DNA MTase enzyme have been regarded as a cancer biomarker or a sign of bacterial diseases. We developed a novel colorimetric method to assay M.SssI MTase activity employing peroxidase-like activity of DNA template Ag/Pt NCs without using restriction enzymes. Based on inhibiting the peroxidase reaction that occurred in the TMB-H2O2 system, in the presence of MTase, a highly sensitive and selective colorimetric biosensor was fabricated with a detection limit (LOD) of 0.05 U/mL and a linear range from 0.5 to 10 U/mL. The changes in absorption intensity were monitored to quantify the M.SssI activity. This strategy had a high selectivity over other proteins. Furthermore, it is also demonstrated that this method can be used for the evaluation and screening of inhibitors for DNA MTase.

Keywords: Ag/Pt nanoclusters; Colorimetric detection; DNA methyltransferase; Enzyme mimic.

MeSH terms

  • Biosensing Techniques
  • Colorimetry / economics
  • Colorimetry / methods*
  • Cost-Benefit Analysis
  • DNA / chemistry
  • DNA / metabolism*
  • DNA Modification Methylases / metabolism*
  • Limit of Detection
  • Nanostructures / chemistry*
  • Peroxidases / metabolism*
  • Platinum / chemistry*
  • Silver / chemistry*
  • Spectrometry, X-Ray Emission
  • Spectrophotometry, Ultraviolet

Substances

  • Silver
  • Platinum
  • DNA
  • Peroxidases
  • DNA Modification Methylases