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Electrochemical Assay for the Signal-On Detection of Human DNA Methyltransferase Activity

Muren, Natalie B. and Barton, Jacqueline K. (2013) Electrochemical Assay for the Signal-On Detection of Human DNA Methyltransferase Activity. Journal of the American Chemical Society, 135 (44). pp. 16632-16640. ISSN 0002-7863. PMCID PMC3899840. doi:10.1021/ja4085918.

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Strategies to detect human DNA methyltransferases are needed, given that aberrant methylation by these enzymes is associated with cancer initiation and progression. Here we describe a nonradioactive, antibody-free, electrochemical assay in which methyltransferase activity on DNA-modified electrodes confers protection from restriction for signal-on detection. We implement this assay with a multiplexed chip platform and show robust detection of both bacterial (SssI) and human (Dnmt1) methyltransferase activity. Essential to work with human methyltransferases, our unique assay design allows activity measurements on both unmethylated and hemimethylated DNA substrates. We validate this assay by comparison with a conventional radioactive method. The advantages of electrochemistry over radioactivity and fluorescence make this assay an accessible and promising new approach for the sensitive, label-free detection of human methyltransferase activity.

Item Type:Article
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URLURL TypeDescription DOIArticle CentralArticle
Barton, Jacqueline K.0000-0001-9883-1600
Additional Information:© 2013 American Chemical Society. Received: August 19, 2013; Published: October 28, 2013. This work was supported by NIH GM61077. We also thank the Parsons Foundation for a fellowship to NBM. We thank C. Pheeney and P. Bartels for assistance with multiplexed chip fabrication. Multiplexed chip fabrication was completed in the Kavli Nanoscience Institute at Caltech.
Group:Kavli Nanoscience Institute
Funding AgencyGrant Number
Ralph M. Parsons FoundationUNSPECIFIED
Issue or Number:44
PubMed Central ID:PMC3899840
Record Number:CaltechAUTHORS:20131205-093720227
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Official Citation:Electrochemical Assay for the Signal-On Detection of Human DNA Methyltransferase Activity Natalie B. Muren and Jacqueline K. Barton. Journal of the American Chemical Society 2013 135 (44), 16632-16640
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:42852
Deposited By: Tony Diaz
Deposited On:05 Dec 2013 17:44
Last Modified:10 Nov 2021 16:29

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