Extended Data Fig. 5: Vh-Ins-HALQ binding to IR and IGF-1R ectodomains.
From: Symmetric and asymmetric receptor conformation continuum induced by a new insulin
a, NanoDSF monitoring of intrinsic protein fluorescence to determine the thermal conformational stability of IR-ECD (top) or IGF1R-ECD (bottom) in the presence of respective ligands in four-times molar excess. Apo-IR-ECD displays two detectable unfolding transitions Tmlow and Tmhigh at 59.2 °C and 63.2 °C, respectively (Supplementary Table 5). The presence of Vh-Ins-HALQ leads to a decrease in Tmlow to 56.3 °C indicating conformational changes induced by ligand binding similarly to insulin (Tmlow = 54.3 °C). Apo-IGF1R-ECD displays a single transition temperature Tmhigh, while binding to hIGF-I leads to an additional melting transition at 57.4 °C. No significant changes in unfolding transitions were observed for IR-ECD in the presence of hIGF-I or for IGF1R-ECD in the presence of Vh-Ins-HALQ or hIns as compared to the respective ligand-free ectodomains. b, MST with IR-ECD (left) and IGF1R-ECD (right) to determine dissociation constants of binding to respective ligands (Supplementary Table 6; n = 3, error bars show standard deviations).
