Extended Data Figure 6. Primary screening and comparison of allosteric parameters of 13 ogerin analogues at GPR68.

The 13 ogerin analogues (structures in Supplementary Table 9) identified from docking a virtual library of over 600 ogerin derivatives were synthesized (Supplementary Information). Production of cAMP was measured in transiently transfected HEK293-T cells at 10 μM and 5 different pH conditions, (a) pH 8.4; (b) pH 7.9; (c) pH 7.4; (d) pH 7.0; and (e) pH 6.5, to reveal any pH-dependent potentiation activity. Normalized results represent mean ± SEM (n = 8 – 16 measurements). (f) Graphic comparison of the allosteric parameters Logα and Logβ. Proton concentration-responses were carried out in the absence and presence of increasing concentrations of ogerin and its analogues, results were analyzed using a standard allosteric operational model to obtain allosteric parameters. Values represent mean ± SEM (n≥3; See details in Supplementary Table 8).