Cholesterol impacts chemokine CCR5 receptor ligand binding activity

International audienceAbstractBackground/AimsThe chemokine CCR5 receptor is target of maraviroc, a negative allosteric modulator of CCR5 that blocks the HIV protein gp120 from associating with the receptor, thereby inhibiting virus cellular entry. As noted with other GPCR family members, the role of the lipid environment in CCR5 signaling remains obscure and very modestly investigated. Controversial literature on the impact of cholesterol depletion in HIV infection and CCR5 signaling, including the hypothesis that cholesterol depletion could inhibit HIV infection, lead us to focus on the understanding of cholesterol impact in the first stages of receptor activation.MethodsTo address this aim, the approach chosen was to employ reconstituted model lipid systems of controlled lipid composition containing CCR5 from two distinct expression systems: P. pastoris and cell‐free expression. The characterization of receptor/ligand interaction in terms of total binding or competition binding assays was independently performed by Plasmon Waveguide Resonance and Fluorescence Anisotropy, respectively. Maraviroc, a potent receptor antagonist was the ligand investigated. Additionally, Coarse‐grained Molecular Dynamics Simulation was employed to investigate cholesterol impact in the receptor conformational flexibility and dynamics.ResultsResults obtained with receptor produced by different expression systems and using different biophysical approaches clearly demonstrate a considerable impact of cholesterol in the binding affinity of maraviroc to the receptor and receptor conformational dynamics. Conclusion: Cholesterol considerably decreases maraviroc binding affinity to the CCR5 receptor. The mechanisms by which this effect occurs seems to involve the adoption of distinct receptor conformational states with restrained structural dynamics and helical motions in the presence of cholesterol