Results
Title

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

Authors

Kermani, Hanie Ahmadzade; Hosseini, Morteza; Miti, Andrea; Dadmehr, Mehdi; Zuccheri, Giampaolo; Hosseinkhani, Saman; Ganjali, Mohammad Reza

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.

Subjects

DNA methyltransferases; PEROXIDASE; SILVER nanoparticles; DNA methylation; GENE expression; BIOSENSORS

Publication

Analytical & Bioanalytical Chemistry, 2018, Vol 410, Issue 20, p4943

ISSN

1618-2642

Publication type

Academic Journal

DOI

10.1007/s00216-018-1143-2