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. 2016 Nov 15;8(1):22–26. doi: 10.1021/acsmedchemlett.6b00395

Figure 5.

Figure 5

Peptide 2 binds irreversibly to Bcl2A1. Fluorescence polarization (FP) measured for Bcl2A1 (A1) binding to fluorescent Bid (FBID) in competition with 1 (no electrophile, black) or 2 (electrophile, green). (A) Kinetics for inhibiting formation of Bcl2A1-FBID complex measured by FP. FBID (5 nM) was saturated with Bcl2A1 (250 nM) before adding peptide 1 or 2 (500 nM) or buffer. (B) Competitive FP assay showing inhibition of Bcl2A1 (15 nM)-FBID (5 nM) complex by increasing concentrations of 2 or 1 after 2 h incubation. (C,D) Titration binding curves of FBID (5 nM) by Bcl2A1 pretreated with 1 (C) or 2 (D) at different protein/peptide ratios. Addition of buffer instead of peptide was used as control (gray, C,D).