Extended Data Fig. 6: Structural features of RO-6889450 and (S)-AMPH bound.

a, 2D representation of interaction of RO-6889450 with hTAAR1. b, Representative dose response curves of the RO-6889450-induced FlAsH-BRET ratio for the mutants of residues Phe154^4.56 and Ser194^5.42 in pocket 3. Values are means ± SEM from three independent experiments performed in triplicate. c, Mutational effects of Phe154^4.56A and Ser194^5.42A on hTAAR1-Gs (left panel), hTAAR1-Gi (middle panel) or hTAAR1-Gq (right panel) signalling in response to RO-6889450 stimulation by Gαs-γ1, Gαi1-γ2 or Gαq-γ9 dissociation assay. Data are presented as mean ± SEM of three independent experiments performed in triplicate. WT, wild type. d, Structural comparison of (S)-AMPH with SEP-363856-bound in hTAAR1 at pocket 1 and pocket 2. e, Conformational changes of key residues from pocket 2 in TAAR1 binds to (S)-AMPH or T1AM. f, The orientation of (S)-AMPH in 500 ns molecular dynamic (MD) simulation course. g, Heatmap was visualized the effects of mutations of key residues from hTAAR1 in response to (S)-AMPH. The heatmap is coloured according to the value of ΔpEC₅₀ (ΔpEC₅₀ = pEC₅₀ of mutant - pEC₅₀ of WT) and Emax (relative WT%). ND, not detected or cannot be established over the tested concentration range. Data are presented as mean ± SEM of three independent experiments performed in triplicate. WT, wild type. h, Representative curve for effects of mutations in pocket 1 (upper panel) and pocket 2 (lower panel) on (S)-AMPH-induced Gs signalling using Gαs-γ1 dissociation assay. Data are presented as mean ± SEM of three independent experiments performed in triplicate. WT, wild type.