Fig. 5: Ligand binding at Y2R NT variants. | Nature Communications

Fig. 5: Ligand binding at Y2R NT variants.

From: Transient ligand contacts of the intrinsically disordered N-terminus of neuropeptide Y2 receptor regulate arrestin-3 recruitment

Fig. 5

a Equilibrium binding between K18-Tamra-NPY and Nluc-Y2R-eYFP variants. Charge reversal in the acidic patches retains wild-type-like equilibrium binding properties, while D42A-S43A-T44A displays a significantly smaller BRET window and about three-fold reduced low-affinity binding compared to wild-type Y2R. b N-terminal deletion in Δ2–41 Y2R increases overall BRET window as expected from the reduced distance, and the low-affinity state has about two-fold reduced affinity compared to wild-type Y2R. Please note that the Y-axis scaling is different between panels (a and b). c Ligand dissociation from the low-affinity state (bound to 300 nM K18-Tamra-NPY; re-binding blocked by 50 µM antagonist) is faster for Δ2–41 and acidic patch charge reversal mutants. Data are the mean ± SEM of n = 3-4 (a, b) or n = 4 (c) independent experiments, performed in technical duplicate or triplicate. * P < 0.05 in one-way ANOVA, with Dunnett’s post-hoc test corrected for multiple comparisons against wild type Y2R. Related to Supplementary Fig. 11: Expression and activity of Nluc-Y2R-eYFP variants used for NanoBRET binding.

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