Ability of the fluorescent probe to monitor dimerization of DAPK catalytic core.
A) Chemical structure of CHPO 187-3 H11-para, which was selected as best hit in the primary and secondary screening assays (absorption and fluorescent spectra of the probe are provided in Supporting Information, Figure S1). B) Binding of CHPO 187-3 H11-para (0.1 µM) to DAPKwt at different concentrations of NH4Ac (• 5 mM, ▪ 100 mM and ▴ 250 mM). Fluorescence polarization (mFP) is plotted versus protein concentration. mFP corresponds to the polarization degree of the fluorescent probe ×103. Error bars indicate the standard deviation of four independent titrations. C) Same than in B) but with DAPKdel.