Identification of novel TAS2R16 agonists. (a) Salicin and 12 related compounds were screened at 10 mM (the highest level practical) for their ability to signal through TAS2R16. The compounds assayed were: 1, salicin; 2, phenyl-β-D-thioglucopyranoside; 3, phenyl-β-D-glucoside; 4, 4-nitrophenyl-β-D-mannopyranoside; 5, hexyl-β-D-glucoside; 6, phenyl-N-acetyl-β-D-glucosaminide; 7, sinigrin; 8, 2-naphthyl-β-D-glucopyranoside; 9, esculin; 10, methyl-β-D-glucopyranoside; 11, phenyl-α-D-glucopyranoside; 12, 1-O-phenyl-β-D-xyloside; 13, phenyl-β-D-galactopyranoside. (b) The structures of salicin and the three TAS2R16 agonists selected for further study. (c) Representative Ca2+ flux traces after addition of salicin or selected analogs. (d) Dose-response curves for salicin and selected analogs. The values for each ligand were normalized to the maximum % over baseline signal (defined as 100%), to highlight the differences in EC50s among the ligands. The EC50 of salicin was 1.2 mM, consistent with its reported value of 3 mM4. Error bars represent the standard deviation, n = 4–8 replicate points.