title: 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.
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, USA.
authors.forename: Wenqing
authors.initials: W
authors.lastname: Xu
authors.forename: Jessica D
authors.initials: JD
authors.lastname: Podoll
authors.forename: Xuan
authors.initials: X
authors.lastname: Dong
authors.forename: Anthony
authors.initials: A
authors.lastname: Tumber
authors.forename: Udo
authors.initials: U
authors.lastname: Oppermann
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
mesh_terms: D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D056466:Histone Demethylases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D015203:Reproducibility of Results
chemical_list: D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D056466:Histone Demethylases