Extended Data Fig. 9: The coupling of ADGRG2-β and ADGRG4-β to G_s.

a, Structural superposition of ADGRG2-β–G_s, ADGRG4-β–G_s and ADGRG3–G_o (PDB: 7D77) complexes when the TM3 helixes of corresponding receptors were aligned. The orientations of α5 helix of G_s rotated by approximately 9° towards the ADGRG2/ADGRG4 compared to the α5 helix of G_o in ADGRG3–G_o complex. b, Comparison of the residues between ADGRG2-β, ADGRG4-β and ADGRG3 that contact with their corresponding downstream G proteins (G_s or G_o) in cryo-EM structures of ADGRG2-β–G_s, ADGRG4-β–G_s and ADGRG3–G_o (PDB: 7D77) complexes, respectively. Contacting residues in ADGRG2-β, ADGRG4-β and ADGRG3 are shown in blue, green and pink dots, respectively, and residues with no interactions are indicated as dashes. Residue positions are labelled with Wootten’s numbers as superscripts on the top of the alignment. Note that ADGRG2 formed more contacts with G_s in the TM2-TM3 region, but the interface showed fewer interactions in the intracellular loops (ICLs) than the interface of GPR97 and G_o. c, d, Detailed interactions between Y391 (c) or L393 (d) of the α5 helix of Gα_s and ADGRG2-β (left panel) or ADGRG4-β (right panel). ADGRG2-β is shown in green, ADGRG4-β in blue. Hydrogen bonds are depicted as red dashed lines. Note that the side chain of L393 of G_s engaged in hydrophobic packing with V800/V2916^5.57, L804/L2920^5.61 and L830/L8945^6.45 of ADGRG2/ADGRG4, respectively. e, f, Detailed interactions between VFNxY motif in ICL2 of ADGRG2-β (e) or ADGRG4-β (f) and G_s. ADGRG2-β is shown in green, ADGRG4-β in blue, respectively. These interactions included hydrogen bonds, cation-π interactions and extended hydrophobic contacts between the VFNxY^ICL2 motif of ADGRG2/ADGRG4 and the residues of the αN helix, β1 strand and β3 strand of G_s. Hydrogen bonds are depicted as red dashed lines.