Extended Data Fig. 3: TAS2R14-Gi-scFv16 and 28.1150µM-TAS2R14-Gg-scFv16 sample preparation, cryo-EM data processing and structure determination.
From: Bitter taste TAS2R14 activation by intracellular tastants and cholesterol
(a, f) Representative size exclusion chromatography (SEC) profiles, SDS-PAGE analysis, representative cryo-EM micrography (scale bar: 50 nm) and selected 2D classification averages of TAS2R14-Gi-scFv16 (a) and 28.1150µM-TAS2R14-Gg-scFv16 (f) complexes. (b, g) Cryo-EM data processing flowcharts of TAS2R14-Gi-scFv16 (b) and 28.1150µM-TAS2R14-Gg-scFv16 (g) complexes by cryoSPARC 4.3. (c, h) Cryo-EM maps are colored by local resolution (Å) for TAS2R14-Gi-scFv16 (c) and 28.1150µM-TAS2R14-Gg-scFv16 (h) complexes. (d, i) Angular distribution of the particles used for final reconstruction (up) and ‘Gold-standard’ FSC curve at the FSC = 0.143 (down) for TAS2R14-Gi-scFv16 (d) and 28.1150µM-TAS2R14-Gg-scFv16 (i) complexes. (e, j) The density maps for helices TM1-TM7 of transmembrane domain, helix 8 and α5 helices of Gα for TAS2R14-Gi-scFv16 (e) and 28.1150µM-TAS2R14-Gg-scFv16 (j) complexes. The SEC and SDS-PAGE experiments were repeated at least three times with similar results.