Figure 1.
High-throughput screening of DAD2 inhibitors using the DSF assay. A, experimental melting curves (top) and derivatives of the melting curves (bottom) obtained for DAD2 in the presence of DMSO, tolfenamic acid, mefenamic acid, and flufenamic acid. Dashed lines, measured melting temperatures of DAD2 from which melting shifts (ΔTm = Tm(compound) − Tm(DMSO)) were calculated. B, chemical structures of the three identified inhibitors, tolfenamic acid, mefenamic acid, flufenamic acid, and of the parent compound, N-phenylanthranilic acid. The melting temperature shift of DAD2 (ΔTm) in the presence of inhibitors is indicated. Ring labels (X and Y) used throughout are labeled on the N-phenylanthranilic acid structure. C, hydrolysis reaction of the synthetic strigolactone GR24 by DAD2/D14 proteins.