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. Author manuscript; available in PMC: 2024 Dec 15.
Published in final edited form as: J Mol Biol. 2023 Nov 2;435(24):168342. doi: 10.1016/j.jmb.2023.168342

Figure 2. Single-molecule FRET analysis of CP α44β84 binding to WASHCAP and V-1. A.

Figure 2.

Transfer efficiency histograms for free CP (0.07 nM Apo-CP, 0 nM ligand, green) and CP titrated with WASHCAP CPI-motif peptide (blue) or V-1 (red). The concentration of CP is low to ensure measurement of single molecules in the confocal volume. WASHCAP induces a shift to a higher Ē value, and V-1 causes a shift to a lower Ē value. Measurements were performed in triplicate; representative histograms are shown. B. Titration curves for WASHCAP (blue) or V-1 (red) plot the mean normalized change in Ē (Ē0 - Ē[WASH/V-1]) vs increasing concentrations of WASHCAP or V-1. Plotted data points are mean values, and error bars are standard deviation. Gray shading represents the 95% confidence interval of the fit as determined by Mathematica.