Extended Data Figure 2. Biochemical preparation of OR51E2-G^s complex bound to propionate.

a) Schematic outlining the strategy for stabilization and purification of the activated OR51E2-G_s complex bound to propionate. b) GloSensor cAMP assay demonstrating that fusion of miniG_s to OR51E2 blocks activation of endogenous G_s in response to treatment with propionate, suggesting that miniG_s couples to the OR51E2 transmembrane core. Data points are the mean of analytical replicates from a representative experiment. Error bars represent the standard deviation between replicates (n=4). c) Size-exclusion chromatogram of purified OR51E2-G_s-Nb35 complex used for structure determinations shown together with a representative SDS-PAGE gel analysis of the collected fraction containing the OR51E2-G_s-Nb35 complex. We observe two bands for OR51E2, likely due to heterogeneous glycosylation of the receptor N-terminus.