A reader-based assay for m6A writers and erasers
| Title | A reader-based assay for m6A writers and erasers |
| Publication Type | Journal Article |
| Year of Publication | 2019 |
| Authors | Wiedmer L., Eberle S.A, Bedi R.K, Śledź P., Caflisch A. |
| Journal | Analytical Chemistry |
| Volume | 91 |
| Issue | 4 |
| Pagination | 3078-3084 |
| Date Published | 2019 Feb 19 |
| Type of Article | Research Article |
| Keywords | DNA methylation, enzyme activity, Enzymes, epitranscriptomics, Fluorescence Resonance Energy Transfer, FRET, High-Throughput Screening Assays, m6A |
| Abstract | We have developed a homogeneous time-resolved fluorescence (HTRF)-based enzyme assay to measure the catalytic activity of N6-methyladenosine (m6A) methyltransferases and demethylases. The assay detects m6A modifications using the natural m6A-binding proteins (m6A readers). The reaction product or substrate m6A-containing RNA and the m6A reader protein are fluorescently labeled such that their proximity during binding initiates Förster resonance energy transfer (FRET). We show that our HTRF assay can be used for high-throughput screening, which will facilitate the discovery of small-molecule modulators of m6A (de)methylases. |
| DOI | 10.1021/acs.analchem.8b05500 |
| pubindex | 0244 |
| Alternate Journal | Anal. Chem. |
| PubMed ID | 30715855 |
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