Figure 6.

Calculating A_2AR–cholesterol binding energetics (A) view of A_2AR bound to cholesterol, in one of several possible binding sites. Colours as in Figure 3A. (B) WTMetaD 2D energy landscape for A_2AR and cholesterol, for the extracellular leaflet only. Three binding regions in red can be seen around the protein, reporting ΔΔG_bind values as follow: S1 = −8 ± 3, S2 = −9 ± 3 and S3 = −5 ± 2 kJ mol^–1. (C) ABFE analysis the sites from panel B. Here, the bound cholesterol is fully decoupled from a POPC membrane whilst bound to A_2AR or whilst free in the membrane. The cycle represented here is a simplified version of the cycle in Supporting Figure 3. The final ΔΔG_bind values are −5 ± 1, −8 ± 1 and −2 ± 1 kJ mol^–1.