Extended Data Fig. 9: Activation of TAAR1 by diverse agonists.

a, Cut-away view of the ligand-binding pocket in the hTAAR1 complexes: T1AM, SEP-363856, (S)-AMPH, RO-6889450, fenoldopam and A77636. b, Cut-away view of the ligand-binding pocket in the mTAAR1 complexes: T1AM, SEP-363856. c, The residues of the core ligand binding pocket are resembled in the SEP-363856-, RO-6889450- and fenoldopam-bound structures. The hydrogen bonds shown as red dashed lines. d, e, Structural representation of the detailed interactions of SEP-363856 and RO-6889450 with the key residues in pocket 3 (e), and SEP-363856, A77636 and fenoldopam with the key residues in pocket 4 (f). The hydrogen bonding is shown as red dashed lines. The red arrows indicate direction of movement. f, Structural superimposition of A77636-bound hTAAR1 with SEP-33856-bound mTAAR1 revealed that Pro183^ECL2 and Tyr287^7.39 in mTAAR1 are different from the allelic residue in hTAAR1. g, Representative curve for effects of the V184^ECL2A and V184^ECL2P mutations in hTAAR1 on A77636-induced Gs signalling using CAMYEL assay. Data are presented as mean ± SEM of three independent experiments performed in triplicate. WT, wild type. h, j, Schematic diagram representing the key residues that contribute to the Gs (h) or Gi (j) proteins coupling with TAAR1. hTAAR1 is depicted as a green cartoon. Residues crucial for Gs or Gi activation are denoted with blue circle (h) or pink circle (j), respectively. i, k, Heatmap illustrating the effect of mutations on the signalling pathways of hTAAR1-Gs (i) and hTAAR1-Gi (k) in response to (S)-AMPH, as evaluated by the Gαs-γ1 and Gαi1-γ2 dissociation assays. Data are presented as mean ± SEM of three independent experiments performed in double.