Fig. 5 |. AS408 uses E122^3.41 of β₂AR, a residue that participates in an allosteric network.

a–k, NAM activity of AS408: β₂AR (wt) (a,d,h), is diminished in E122Q (b,e,i), E122L (c,f,j) and in E122R (g,k), in norepinephrine-stimulated β-arrestin 2 recruitment (HEK293 cells), [³⁵S]GTPγS binding (Sf9 cells) (a–c) and cAMP accumulation (HEK293 cells) (d–g), compared to β₂AR (wt) (h–k). l, β₂AR (E122R) displayed a higher basal activity but was unresponsive to inverse agonist ICI-118,551 (CHO cells). Expression of β₂AR (wt) and E122R in l were measured by [³H]DHAP binding assays. [³⁵S]GTPγS binding assays were performed on membranes prepared from Sf9 cells coexpressing β₂AR (or mutant) and Gsαβγ. Data are derived from 2–12 experiments done in duplicate and are given as mean ± s.e.m. if n ≥ 3, or as mean if n = 2. The sample size (n) is labeled in the figure.