Figure 4 |. MIPS521 stabilises the A1R-Gi2 ternary complex.

a-d, RMSD (Å) of ADO from GaMD simulations completed in the (a) absence or (b) presence of MIPS521, (c) G_i2, or (d) both G_i2 and MIPS521. e-h, Distance between the intracellular ends of TM3 and TM6 (measured as the distance in Å between Arg105^3.50 and Glu229^6.30) in the (e) absence or (f) presence of MIPS521, (g) G_i2, or (h) both G_i2 and MIPS521. Each condition represents three GaMD simulations, with each simulation trace displayed in a different colour (black, red, blue). The lines depict the running average over 2 ns. i, MIPS521 has no effect on the dissociation rate of [³H]DPCPX, promoted by isotopic dilution with excess antagonist, SLV320 (1 μM; Control), at the A1R alone reconstituted in rHDL. Addition of a saturating concentration of G_i results in a retardation of [³H]DPCPX dissociation that is reduced by co-addition of MIPS521. Data is mean ± SEM, n = 3-5 j, Quantification of dissociation rates from traces. Data is mean ± SEM, n ≥ 3 experiments performed in duplicate. *P < 0.05 (compared to control; one-way ANOVA, Tukey’s post hoc test). #P < 0.05 (compared to control in the presence of G_i2; one-way ANOVA, Tukey’s post hoc test).