Extended Data Fig. 6: Possible Lipid binding sites and phosphorylation sites.

a, Gβγ subunits was modeled into the NTSR1–GRK2 structure by structural alignment of GRK2 from NTSR1–GRK2 structure and GRK2–Gαq–Gβγ structure complex (PDB code: 2BCJ). GRK2 was shown in blue and GRK5 (PDB code: 6PJX) was shown in green. The possible lipid binding sites from GRK2 are highlighted in orange and GRK5 are highlighted in dark red. NLBD, the N-terminal lipid binding site. CLBD, the C-terminal lipid binding site. b, WT and mutated GRK2 recruitment to NTSR1 induced by NTS (left panel), relative expression level of WT and mutated GRK2 (right panel). NLBD refers to the combined mutations of N-terminal basic residues (R27A, K30A, K31A). CLBD means the combined mutations R535A, R539A, K540A, K541A, K543A, K545A. Combined mutation refers to all the mutations combined. Data were shown as mean ± S.D. from three independent experiments (n = 3), performed in duplicate. Statistical significance of differences between WT and mutants was determined by two-sided one-way ANOVA. *P < 0.05 and **P < 0.01 and ***P < 0.001 versus WT. The detailed information is provided in Extended Data Table 3. c, Phosphorylation sites of the active NTSR1. The extended loop of ICL3 or the elongated C-terminal tail of NTSR1 could reach the active cleft of GRK2, thus are available to be phosphorylated by GRK2. P represents phosphorylation modifications.