A Reader-Based Assay for m6A Writers and Erasers

Autor:	L. Wiedmer, R.K. Bedi, S.A. Eberle, Paweł Śledź, Amedeo Caflisch
Rok vydání:	2019
Předmět:	Förster resonance energy transfer Methyltransferase Biochemistry Chemistry Homogeneous 010401 analytical chemistry RNA 010402 general chemistry 7. Clean energy 01 natural sciences 0104 chemical sciences Analytical Chemistry Reaction product
Zdroj:	Analytical Chemistry. 91:3078-3084
ISSN:	1520-6882 0003-2700
DOI:	10.1021/acs.analchem.8b05500
Popis:	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 Forster 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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::5afd3a164a53fafdb135dba24743a59d https://doi.org/10.1021/acs.analchem.8b05500 Zobrazit plný text záznamu