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. 2016 Feb 15;129(4):831–842. doi: 10.1242/jcs.180182

Fig. 2.

Fig. 2.

TASK-1 C-terminus binds to 14-3-3 proteins with 10- to 100-fold lower affinity than the TASK-3 C-terminus. Fluorescence polarization titration of fluorescein-labeled TASK-1 pS393 (black), TASK-3 pS373 (red) and TASK-3 pS373 ΔV374 (blue) peptide (10 nM) with increasing concentrations of the indicated 14-3-3 isoform. Fluorescein was excited at 485 nm, and fluorescence polarization was monitored at 535 nm. A monophasic fit was used to fit the data (Goodness-of-fit: R2 from 0.76 to 0.98). Equilibrium constants determined from the fit are listed in Table 1. No binding was detectable for the unphosphorylated TASK C-terminal peptides. The assay was repeated at least six times for each data point with two batches of independently purified 14-3-3 proteins. Error bars depict s.e.m. (A) 14-3-3β, (B) 14-3-3γ, (C) 14-3-3ε, (D) 14-3-3ζ, (E) 14-3-3η and (F) 14-3-3τ.