Abstract
We previously reported methylstat as a selective inhibitor of jumonji C domain-containing histone demethylases (JHDMs). Herein, we describe the synthesis of a fluorescent analogue of methylstat and its application as a tracer in fluorescence polarization assays. Using this format, we have evaluated the binding affinities of several known JHDM probes, as well as the native cofactor and substrate of JHDM1A. This fluorophore allowed a highly robust and miniaturized competition assay sufficient for high-throughput screening.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Enzyme Inhibitors / chemical synthesis
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Enzyme Inhibitors / chemistry*
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Enzyme Inhibitors / pharmacology
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Fluorescence Polarization
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Fluorescent Dyes / chemical synthesis
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Fluorescent Dyes / chemistry*
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Fluorescent Dyes / pharmacology
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Histone Demethylases / antagonists & inhibitors
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Histone Demethylases / chemistry
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Histone Demethylases / metabolism*
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Reproducibility of Results
Substances
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Enzyme Inhibitors
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Fluorescent Dyes
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Histone Demethylases