Extended Data Fig. 9: Key residues and ligand structures of the biased agonism.
From: Structural basis of sphingosine-1-phosphate receptor 1 activation and biased agonism
a, Flow cytometry-based surface expression analysis for the mutants studied in the β-arrestin assays. The vertical dashed line separates mutants that are included in Fig. 4c (left) and Extended Data Fig. 8b (right). Note that the expression data of the W2696.48A mutant is identical to the one shown in Extended Data Fig. 5c. Data are presented as mean values ± SEM, n = 3-5 (dots). b, Mutant study to assess S1P response of the indicated mutants by the NanoBiT-Gi dissociation assay and the β-arrestin recruitment assay. Dashed lines and the long dashed dotted lines in the mutant panels represent the WT and the mock responses, respectively. Data are presented as mean values ± SEM, n = 3-4. Note that, in many data points, error bars are smaller than the size of symbols and thus are not visible. c, Bias profiles of etrasimod and S1P receptor agonist 1. Gi dissociation, β-arrestin recruitment and β-arrestin endosomal translocation responses in the wild-type S1PR1 were tested by the NanoBiT-based assays. For the individual experiments performed in parallel, data were normalized to S1P and presented as a logarithm of relative intrinsic activity (RAi; EC50/Emax value relative to that of S1P; Methods). Data are presented as mean values ± SEM, n = 3-4.
