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Quantitative analysis of histone demethylase probes using fluorescence polarization.

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.

SUBMITTER: Xu W

PROVIDER: S-EPMC3724763 | biostudies-other | 2013 Jun

REPOSITORIES: biostudies-other

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Quantitative analysis of histone demethylase probes using fluorescence polarization.

Xu Wenqing W Podoll Jessica D JD Dong Xuan X Tumber Anthony A Oppermann Udo U Wang Xiang X

Journal of medicinal chemistry 20130613 12

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 ...[more]

PMID: 23725560

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