Fig. 1: Cryo-EM DP1 structures in the inactive and active G_s protein bound states.

a Cryo-EM density of the inactive state DP1_ICL3bRIL construct (DP1—blue, bRIL—yellow) bound to the inverse agonist ONO3030297 (cyan) and in complex with the anti-bRIL Fab (BAG2, heavy chain—orange, light chain—pink) and stabilizing nanobody (Nb, light green). The insert shows EM densities in the ligand binding pocket for ONO3030297, ONO2550289, and APO receptor. b Cryo-EM density of DP1 (salmon) bound to PGD2 (gold) and in complex with heterotrimeric G_s protein (Gα_s—cyan, Gβ—sand, Gγ—violet) and nanobody Nb-35 (green). The insert shows EM densities in the ligand binding pocket for agonists PGD2 and BW245C. The ligands in (a–c) are shown as sticks with carbon atoms colored in cyan (ONO3030297), chartreuse (ONO2550289), gold (PGD2) or violet (BW245C). c Structures of DP1 in the inactive state and in the active Gs protein-bound states. Two gray lines indicate membrane boundaries as provided by the Orientation of Proteins in Membranes (OPM) database. d Cellular cAMP production assay with transiently transfected WT DP1. The agonist data are normalized to the basal signal (set at 0%) and the maximal PGD2-induced signal (set at 100%). The inverse agonist data are normalized to the basal signal (set at 0%) and the signal from the empty vector-transfected cells (set at −100%). Data points correspond to means ± SEM for three biologically independent experiments conducted in triplicates. The corresponding E_max and EC₅₀ values are shown in Supplementary Table 3. Source Data are provided as a Source data file.