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Fig. 4 Figure 4. The biophysical and biochemical characterization of DC-TEAD3in03. (A) The chemical structures of DC-TEAD3in01, DC-TEAD3in02 and DC-TEAD3in03. (B)‒(D) The inhibitory activities of DC-TEAD3in01, DC-TEAD3in02 and DC-TEAD3in03 through CuAAC-based click chemistry. The band intensity was quantified in ImageJ (NIH). The experiments were performed in triplicate and the data was shown as mean ± SD, n = 3. (E) DC-TEAD3in03 selectively inhibited TEAD1 and TEAD3 palmitoylation while sparing TEAD2 and TEAD4. (F) and (G) DC-TEAD3in03 significantly improved the thermostability of TEAD3 but had minimal effect on TEAD1 or corresponding C359S mutant. Synthetic YAP peptide was used as the positive control with the ratio of 1:5. The experiments were performed in triplicate. (H) The modification of transfected-TEAD3 upon 50 μmol/L DC-TEAD3in03 treatment for 6 h was detected by MS experiments.