A Colorimetric Assay of Dna Methyltransferase Activity Based on Peroxidase Mimicking of Dna Template Ag/Pt Bimetallic Nanoclusters Publisher Pubmed



Kermani HA¹ ; Hosseini M¹ ; Miti A² ; Dadmehr M³ ; Zuccheri G² ; Hosseinkhani S⁴ ; Ganjali MR^{5, 6}
Source: Analytical and Bioanalytical Chemistry Published:2018

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. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature.