Putative cholesterol‐binding sites in human immunodeficiency virus (HIV) coreceptors CXCR4 and CCR5

Zhukovsky, Mikhail A.; Ott, Albrecht; Helms, Volkhard

doi:10.1002/prot.24211

Cited by 21 publications

(22 citation statements)

References 82 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, enhanced binding to possible cholesterol recognition amino acid consensus (CRAC) motifs, as suggested for the serotonin 5-hydroxytryptamine(1A) receptor [44], did not appear in the simulations, eventhough three CRAC motifs could be identified in the sequence of CXCR4. These observations are in agreement with a cholesterol docking study on CXCR4 [45]. …”

Section: Resultssupporting

confidence: 92%

Dynamic Cholesterol-Conditioned Dimerization of the G Protein Coupled Chemokine Receptor Type 4

et al. 2016

View full text Add to dashboard Cite

G protein coupled receptors (GPCRs) allow for the transmission of signals across biological membranes. For a number of GPCRs, this signaling was shown to be coupled to prior dimerization of the receptor. The chemokine receptor type 4 (CXCR4) was reported before to form dimers and their functionality was shown to depend on membrane cholesterol. Here, we address the dimerization pattern of CXCR4 in pure phospholipid bilayers and in cholesterol-rich membranes. Using ensembles of molecular dynamics simulations, we show that CXCR4 dimerizes promiscuously in phospholipid membranes. Addition of cholesterol dramatically affects the dimerization pattern: cholesterol binding largely abolishes the preferred dimer motif observed for pure phospholipid bilayers formed mainly by transmembrane helices 1 and 7 (TM1/TM5-7) at the dimer interface. In turn, the symmetric TM3,4/TM3,4 interface is enabled first by intercalating cholesterol molecules. These data provide a molecular basis for the modulation of GPCR activity by its lipid environment.

show abstract

Section: Resultssupporting

confidence: 92%

Dynamic Cholesterol-Conditioned Dimerization of the G Protein Coupled Chemokine Receptor Type 4

et al. 2016

View full text Add to dashboard Cite

show abstract

“…A, TM helices VII, V, VI, IV, and even I are involved in the Chol–protein interactions. These helices have been identified as Chol occupancy sites in others GPCRs . TM segments already reported as Chol binding sites were also found as highly probable sites all along the course of our simulations.…”

Section: Resultssupporting

confidence: 71%

Cholesterol impacts chemokine CCR5 receptor ligand‐binding activity

Calmet

Cullin

Cortès³

et al. 2019

The FEBS Journal

View full text Add to dashboard Cite

The 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 G‐protein‐coupled receptor family members, the role of the lipid environment in CCR5 signaling remains obscure and very modestly investigated. Controversial literature on the impact of cholesterol (Chol) depletion in HIV infection and CCR5 signaling, including the hypothesis that Chol depletion could inhibit HIV infection, lead us to focus on the understanding of Chol impact in the first stages of receptor activation. To address this aim, the approach chosen was to employ reconstituted model lipid systems of controlled lipid composition containing CCR5 from two distinct expression systems: Pichia 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 Chol impact in the receptor‐conformational flexibility and dynamics. Results obtained with receptor produced by different expression systems and using different biophysical approaches clearly demonstrate a considerable impact of Chol in the binding affinity of maraviroc to the receptor and receptor‐conformational dynamics. Chol considerably decreases maraviroc binding affinity to the CCR5 receptor. The mechanisms by which this effect occurs seem to involve the adoption of distinct receptor‐conformational states with restrained structural dynamics and helical motions in the presence of Chol.

show abstract

“…Yet, an aspect that remains unanswered is whether cholesterol effect in the antagonist-induced receptor conformational change is a pure result of the alterations in the physical properties of the membrane (e.g., fluidity) or by a direct binding to the CCR5 receptor or a combination of both. Indeed, a putative-cholesterol binding site has been identified in human CXCR4 by docking analysis and in CCR5 by sequence alignment53 and cholesterol was even used in CXCR4 crystallization54.…”

Section: Discussionmentioning

confidence: 99%

Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids

Calmet

Maria

Harté

et al. 2016

Sci Rep

View full text Add to dashboard Cite

G-protein coupled receptors (GPCRs) are important therapeutic targets since more than 40% of the drugs on the market exert their action through these proteins. To decipher the molecular mechanisms of activation and signaling, GPCRs often need to be isolated and reconstituted from a detergent-solubilized state into a well-defined and controllable lipid model system. Several methods exist to reconstitute membrane proteins in lipid systems but usually the reconstitution success is tested at the end of the experiment and often by an additional and indirect method. Irrespective of the method used, the reconstitution process is often an intractable and time-consuming trial-and-error procedure. Herein, we present a method that allows directly monitoring the reconstitution of GPCRs in model planar lipid membranes. Plasmon waveguide resonance (PWR) allows following GPCR lipid reconstitution process without any labeling and with high sensitivity. Additionally, the method is ideal to probe the lipid effect on receptor ligand binding as demonstrated by antagonist binding to the chemokine CCR5 receptor.

show abstract

Putative cholesterol‐binding sites in human immunodeficiency virus (HIV) coreceptors CXCR4 and CCR5

Cited by 21 publications

References 82 publications

Dynamic Cholesterol-Conditioned Dimerization of the G Protein Coupled Chemokine Receptor Type 4

Dynamic Cholesterol-Conditioned Dimerization of the G Protein Coupled Chemokine Receptor Type 4

Cholesterol impacts chemokine CCR5 receptor ligand‐binding activity

Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids

Contact Info

Product

Resources

About