Evidence that specific interactions play a role in the cholesterol sensitivity of G protein-coupled receptors

G protein-coupled receptors (GPCRs) are known to be modulated by membrane cholesterol levels, but whether or not the effects are caused by specific receptor-cholesterol interactions or cholesterol's general effects on the membrane is not well-understood. We performed coarse-grained molecular dynamics (CGMD) simulations coupled with structural bioinformatics approaches on the beta(2)-adrenergic receptor (beta(2)AR) and the cholecystokinin (CCK) receptor subfamily. The beta(2)AR has been shown to be sensitive to membrane cholesterol and cholesterol molecules have been clearly resolved in numerous beta(2)AR crystal structures. The two CCK receptors are highly homologous and preserve similar cholesterol recognition motifs but despite their homology, CCK1R shows functional sensitivity to membrane cholesterol while CCK2R does not. Our results offer new insights into how cholesterol modulates GPCR function by showing cholesterol interactions with beta(2)AR that agree with previously published data; additionally, we observe differential and specific cholesterol binding in the CCK receptor subfamily while revealing a previously unreported Cholesterol Recognition Amino-acid Consensus (CRAC) sequence that is also conserved across 38% of class A GPCRs. A thermal denaturation assay (LCP-T-m) shows that mutation of a conserved CRAC sequence on TM7 of the beta(2)AR affects cholesterol stabilization of the receptor in a lipid bilayer. The results of this study provide a better understanding of receptor-cholesterol interactions that can contribute to novel and improved therapeutics for a variety of diseases.

Automated summary

This study investigates the modulation of GPCRs by cholesterol through simulations and analysis, revealing specific interactions between cholesterol and beta(2)AR, as well as differential cholesterol binding in the CCK receptor subfamily. The findings contribute to a better understanding of receptor-cholesterol interactions and may lead to the development of novel therapeutics for various diseases.