Journal: Journal of medicinal chemistry

Article Title: Discovery of a Novel Chemotype of Histone Lysine Methyltransferase EHMT1/2 (GLP/G9a) Inhibitors: Rational Design, Synthesis, Biological Evaluation, and Co-crystal Structure

doi: 10.1021/acs.jmedchem.8b02008

Figure Lengend Snippet: Direct binding measurements of the interaction between G9a and compounds 7 (left) and 12a (right). (a, b) MST binding affinity assays were performed as described in Experimental Section. Representative dose–response curves of 7 (a) and 12a (b) binding to G9a are shown; Fnorm, normalized fluorescence. Experiments were repeated independently three times. Reported KD is the mean ± SD of three independent experiments. (c, d) Sensorgrams obtained from the SPR interaction analysis of compounds 7 (c) and 12a (d) binding to immobilized G9a. Each compound was injected at eight different concentrations (1.56, 3.12, 6.25, 12.5, 25, 50, 100, and 200 μM). The equilibrium dissociation constants (KD) were derived from the ratio between kinetic dissociation (koff) and association (kon) constants.

Article Snippet: Pretests using standard-treated and premium-coated MST capillaries (NanoTemper Technologies) were performed to test for adsorption of NT647-G9a to capillary walls by analyzing capillary scans recorded by the Monolith NT.115pico prior to MST experiments.

Techniques: Binding Assay, Fluorescence, Injection, Derivative Assay