Extended Data Fig. 5: Pharmacological characterization of SEP-363856 at TAAR1 and 5-HT1AR.
From: Ligand recognition and G-protein coupling of trace amine receptor TAAR1
a, 2D representation of interaction of SEP-363856 with h/mTAAR1. b, The superimposed structures of SEP-363856-bound hTAAR1 (green/blue) with SEP-363856-bound mTAAR1 (yellow/cyan) reveal nearly identical binding pose of ligand. c, d, Comparison of the binding mode of SEP-363856 and T1AM in hTAAR1 (c) and mTAAR1 (d). e-g, Glosensor assay of receptors WT and mutations in hTAAR1 (e), mTAAR1 (f) and 5-HT1AR (g) on SEP-363856-induced Gs signalling. Data represent mean ± SEM from three independent experiments performed in triplicate. WT, wild type. h, Competitive binding curves of SEP-363856 to wild type hTAAR1 or its mutants in pocket 1 and pocket 2. [3H]-tyramine was used in our study. The Ki values of wild type hTAAR1 for SEP-363856 are 22.59 ± 0.60 nM. Data are presented as mean ± SEM of three independent experiments performed in triplicate. i, j, Representative curve for effects of the Phe186ECL2 mutations in hTAAR1 (i) or Phe185ECL2 mutations in mTAAR1 (j) on SEP-363856-induced Gs signalling detected by Glosensor assay. Data are presented as mean ± SEM of three independent experiments performed in triplicate. WT, wild type. k, The residues in ECL2 of TAAR1 are sequentially aligned among aminergic receptors. * Representing the corresponding residues in aminergic receptors that relatively to Phe186ECL2 of hTAAR1. l, Structural comparison of the hTAAR1-SEP363856 with 5-HT1AR-SEP363856 reveals that the residues Phe186ECL2 in hTAAR1 and Lys191ECL2 in 5-HT1AR are different. Residues are shown as dodger blue sticks in hTAAR1 and pale violet red sticks in 5-HT1AR, respectively. m, Representative curve for effects of the Lys191ECL2A mutation in 5-HT1AR on SEP-363856-induced Gi signalling detected by Glosensor assay. Data are presented as mean ± SEM of three independent experiments performed in triplicate. WT, wild type.
