Extended Data Fig. 7: Intramolecular interaction between IP15 and ADGRG2(FL).
From: Tethered peptide activation mechanism of the adhesion GPCRs ADGRG2 and ADGRG4
The therapeutic capacities of peptides derived from Stachel sequence are worth noting. For example, the GPR126 agonist derived from Stachel sequence was able to rescue the phenotype caused by human GPR126 disease associated mutant in a zebrafish model. In addition, Stachel-derived peptides of GPR116/ADGRF5 has therapeutic potential to modulate endogenous alveolar surfactant pools to treat pulmonary diseases associated with surfactant dysfunction. a, Representative dose-response curves for VPM-p15 and IP15 induced cAMP accumulation in HEK293 cells overexpressing ADGRG2-β(ΔS) (ADGRG2-β without Stachel sequence, residues 614–1009) using a GloSensor assay. Values are the mean ± s.e.m. of three independent experiments (n = 3) performed in triplicate. b, EM densities allowed unambiguous assignment of the side chains of the residues in the lower rim of U shape in IP15 (red). The Stachel sequence of ADGRG2 (blue) in ADGRG2-β–Gs complex was aligned with IP15 as a reference. Note that EM density corresponding to upper rim of the U shape was absent compared with the ADGRG2-β–Gs complex structure, and ten out of thirteen residues of the IP15 peptide could be traced according to the EM density. c, The hydrophobic interactions between the I603 of IP15 (red stick) and surrounding residues (pale green sticks) in ADGRG2(FL). d–i, EM densities of the Stachel sequence residues I603 (c), I599 (d), 4-MeF601 (e), L604 (f), L605 (g), L607 (h) (shown in red) and their surrounding residues (shown in green) of ADGRG2(FL) in IP15-ADGRG2(FL)–Gs complex structure. Corresponding residues in the ADGRG2-β–Gs complex was aligned as a reference, Stachel sequence residues are shown in blue, and the surrounding residues are shown in grey. j, RMSD analysis of 4-MeF601 and F601 in 200 ns trajectories. RMSD of 4-MeF601 and its interacting residues during 200 ns MD simulation (upper panel), and RMSD of F601 and its interaction residues during 200 ns MD simulation (lower panel) are shown. k–r, Effects of mutations in IP15 on cAMP accumulation in ADGRG2-β(ΔS) (ADGRG2-β without Stachel sequence, residues 614–1009) overexpressed HEK293 cells. Values are the mean ± s.e.m. of three independent experiments (n = 3) performed in triplicate.
