title: False HDAC Inhibition by Aurone Compound.
abstract: Fluorescence assays are useful tools for estimating enzymatic activity. Their simplicity and manageability make them suitable for screening enzyme inhibitors in drug discovery studies. However, researchers need to pay attention to compounds that show auto-fluorescence and quench fluorescence, because such compounds lower the accuracy of the fluorescence assay systems by producing false-positive or negative results. In this study, we found that aurone compound 7, which has been reported as a histone deacetylase (HDAC) inhibitor, gave false-positive results. Although compound 7 was identified by an in vitro HDAC fluorescence assay, it did not show HDAC inhibitory activity in a cell-based assay, leading us to suspect its in vitro HDAC inhibitory activity. As a result of verification experiments, we found that compound 7 interferes with the HDAC fluorescence assay by quenching the HDAC fluorescence signal. Our findings underscore the faults of fluorescence assays and call attention to careless interpretation.
authors.affiliation: Graduate School of Medical Science, Kyoto Prefectural University of Medicine.
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Itoh
authors.forename: Miki
authors.initials: M
authors.lastname: Suzuki
authors.forename: Taiji
authors.initials: T
authors.lastname: Matsui
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Ota
authors.forename: Zi
authors.initials: Z
authors.lastname: Hui
authors.forename: Kazunori
authors.initials: K
authors.lastname: Tsubaki
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D016477:Artifacts; D001572:Benzofurans / Q000737:chemistry / Q000494:pharmacology*; D057075:Enzyme Assays / Q000379:methods / Q000592:standards*; D005453:Fluorescence; D006367:HeLa Cells; D056572:Histone Deacetylase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D015203:Reproducibility of Results; D014407:Tumor Cells, Cultured
chemical_list: D001572:Benzofurans; D056572:Histone Deacetylase Inhibitors; C486837:aurone; D006655:Histone Deacetylases