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

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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.

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Acknowledgements

The authors thank the research Council of University of Tehran (Grant 28645/01/02) for financial support of this work.

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Authors and Affiliations

  1. Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Amir Abad, North Kargar St, Tehran, 1417466191, Iran

    Hanie Ahmadzade Kermani & Morteza Hosseini

  2. Department of Pharmacy and Biotechnology and Interdepartmental Center for Industrial Research on Health Sciences and Technologies at the University of Bologna, The Nanoscience Institute of CNR, the National Interuniversity Consortium of Materials Science and Technology, Via Irnerio, 48, 40126, Bologna, Italy

    Andrea Miti & Giampaolo Zuccheri

  3. Department of Biotechnology, Payame Noor University, Lashkarak Rd, P.O. Box 19395-4697, Tehran, Iran

    Mehdi Dadmehr

  4. Department of Biochemistry, Tarbiat Modares University, Jalal AleAhmad Nasr, P.O. Box 14115-111, Tehran, Iran

    Saman Hosseinkhani

  5. Center of Excellence in Electrochemistry, School of Chemistry, College of Science, University of Tehran, 16th Azar St., Enghelab Sq, P.O. Box 14155-6619, Tehran, Iran

    Mohammad Reza Ganjali

  6. Biosensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, No 10 Jalale Al Ahmad St, Tehran, 1411713137, Iran

    Mohammad Reza Ganjali

Authors
  1. Hanie Ahmadzade Kermani
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  2. Morteza Hosseini
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  3. Andrea Miti
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  4. Mehdi Dadmehr
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  6. Saman Hosseinkhani
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  7. Mohammad Reza Ganjali
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Correspondence to Morteza Hosseini.

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Kermani, H.A., Hosseini, M., Miti, A. et al. A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters. Anal Bioanal Chem 410, 4943–4952 (2018). https://doi.org/10.1007/s00216-018-1143-2

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  • DOI: https://doi.org/10.1007/s00216-018-1143-2

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