Extended Data Fig. 8: Interaction mode of AA-14 with GPR101.
From: Structure of GPR101–Gs enables identification of ligands with rejuvenating potential
a, The binding energy contribution of side pocket residues in the AA-14-GPR101-Gs complex calculated by the molecular mechanics Poisson–Boltzmann surface area (MM-PBSA) method. Data are from three independent MD simulations (n = 3). b-c, Cryo-EM density maps of AA-14 and its interacting residues at the side binding pocket (b) and the second binding pocket (c) of GPR101. The density maps were shown with the same counter level set in Chimera software (0.0110 V). d, Schematic representation of the FlAsH-BRET assay design. NanoLuc (Nluc) was fused to N-terminus of GPR101, and the FlAsH motif (CCPGCC) was incorporated at the designated positions of the extracellular loops (ECLs) of the receptor. The WT GPR101 and corresponding mutants were expressed at similar levels. e, The maximal response of GPR101 FlAsH-BRET sensors in response to AA-14 stimulation. Values are from three independent experiments (n = 3). ND, not detectable. f, Effects of mutations in the second binding pocket on AA-14-induced ECL1 conformational changes of GPR101 measured by FlAsH-BRET. Values are from three independent experiments (n = 3). The WT GPR101 and corresponding mutants were expressed at similar levels. For a, e and f, data are presented as mean ± SEM values. P values were calculated using one-way ANOVA with Dunnett’s post hoc test and are indicated in the graphs.
