Extended Data Fig. 4: Purification and structure determination of SEP-363856 bound TAAR1-Gs and 5HT1AR-Gi complexes.
From: Recognition of methamphetamine and other amines by trace amine receptor TAAR1
(a) Representative size exclusion chromatography (SEC) profiles and SDS-PAGE analysis of TAAR1-Gs complex activated by SEP-363856. Experiment was repeated at least three times with similar results. (b-c) Representative cryo-EM image from 7,262 movies (b) and 2D classification averages (c) of SEP-363856-TAAR1-Gs. (d) Cryo-EM data processing flowcharts of SEP-363856-TAAR1-Gs by cyroSPARC 3.2. (e) The Fourier shell correlation (FSC) curves of SEP-363856-TAAR1-Gs. The global resolution of the final processed density map estimated at the FSC = 0.143 is 2.6 Å. (f) The global density map of SEP-363856-TAAR1-Gs colored by local resolutions. (g) The density maps of helices TM1–TM7 of transmembrane domain, extracellular loop ECL2 of TAAR1, α5 helix of Gαs, and SEP-363856 in SEP-363856-TAAR1-Gs complex. (h) Representative size exclusion chromatography (SEC) profiles and SDS-PAGE analysis of 5HT1AR -Gi complex activated by SEP-363856. Experiment was repeated at least three times with similar results. (i-j) Representative cryo-EM image from 6,222 movies (i) and 2D classification averages (j) of SEP-363856-5HT1AR -Gi. (k) Cryo-EM data processing flowcharts of SEP-363856-5HT1AR-Gi complex by cyroSPARC 3.2. (l) The Fourier shell correlation (FSC) curves of SEP-363856-5HT1AR -Gi complex. The global resolution of the final processed density map estimated at the FSC = 0.143 is 3.0 Å. (m) The global density map of SEP-363856-5HT1AR-Gi colored by local resolutions. (n) The density maps of helices TM1–TM7 of transmembrane domain, α5 helix of Gαi, and SEP-363856 in 5HT1AR -Gi complex.
