Extended Data Fig. 6: The side binding pocket of GPR101.
From: Structure of GPR101–Gs enables identification of ligands with rejuvenating potential
a, The side pocket volumes of GPR101 (slate) and GPR52 (green). The PyVOL, which is a python library packaged into a PyMOL GUI, was used to calculate the side pocket volumes with the probe radius set at 1.4 Å. b-c, Sequence alignment of c17-binding side pocket residues at the N-terminus (b) and TM bundles or ECL2 (c) in GPR52 with the equivalent residues in GPR101. The conserved residues are shown in blue, while the distinct residues are shown in red. d, Effects of substitution of key residues in the side pocket of GPR101 with the equivalent residues of GPR52 on the responsiveness of GPR101 to c17 stimulation. Data were normalized to the basal cAMP level in HEK293 cells transfected with WT GPR101 and are from three independent experiments (n = 3). The WT GPR101 and corresponding mutants were expressed at similar levels. e, Cell surface ELISA assay showing similar expression levels of WT GPR101, WT GPR52 and GPR101 mutants with key residues in side pocket substituted with the equivalent residues of GPR52. Data were normalized to the expression level of WT GPR101 in HEK293 cells. Data were from three independent experiments (n = 3). For d-e, all data are presented as mean ± SEM values. P values were calculated using two-way ANOVA (d) or one-way ANOVA (e) with Dunnett’s post hoc test and are indicated in the graphs.
