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Updated: Jun 3, 2026

Detection of Modified Forms of Cytosine Using Sensitive Immunohistochemistry

Detection of Modified Forms of Cytosine Using Sensitive Immunohistochemistry

Published on: August 16, 2016

Phosphopeptides designed for 5-methylcytosine recognition.

Akiko Nomura¹, Akimitsu Okamoto

¹Advanced Science Institute, RIKEN, Wako, Saitama 351-0198, Japan.

|March 23, 2011

Summary

Researchers designed a novel phosphopeptide that selectively binds to methylated DNA (5-methylcytosine). This peptide, utilizing zinc finger motifs, can detect genomic DNA methylation status via fluorescence anisotropy in minutes.

Area of Science:

Biochemistry
Molecular Biology
Genetics

Background:

DNA methylation is a crucial epigenetic modification.
Selective binding to methylated DNA is essential for epigenetic research.
Zinc finger motifs are known DNA-binding domains.

Purpose of the Study:

To develop a rationally designed artificial phosphopeptide for selective DNA methylation detection.
To investigate the interaction between phosphopeptides and 5-methylcytosine in DNA.
To establish a rapid method for assessing genomic DNA methylation status.

Main Methods:

Rational design of a phosphopeptide with two tandem zinc finger motifs.
Modification of a zinc finger motif by replacing glutamate with phosphotyrosine.

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Selective Capture of 5-hydroxymethylcytosine from Genomic DNA

Selective Capture of 5-hydroxymethylcytosine from Genomic DNA

Published on: October 5, 2012

DNA binding assays to assess selectivity for 5-methylcytosine.

Fluorescence anisotropy measurements using a fluorescence-labeled phosphopeptide.

Main Results:

The artificial phosphopeptide demonstrated selective binding to 5-methylcytosine in duplex DNA.
The phosphotyrosine residue was key to the methylcytosine selectivity of DNA binding.
Rearrangement of zinc finger peptides allowed flexible modulation of target methylated sequences.
Fluorescence-labeled phosphopeptide detected genomic DNA methylation status within 10 minutes via fluorescence anisotropy.

Conclusions:

A novel phosphopeptide has been engineered for specific recognition of methylated DNA.
This phosphopeptide offers a new tool for studying DNA methylation and epigenetic modifications.
The developed method provides a rapid and sensitive approach for determining DNA methylation status.