Cholesterol is essential for macrophage inflammatory protein 1beta binding and conformational integrity of CC chemokine receptor 5

@article{citeulike:2602803, abstract = {The chemokine receptor, CCR5, is used as a human immunodeficiency virus coreceptor in combination with CD4 during transmission and early infection. CCR5 has been shown to be palmitoylated and targeted to cholesterol- and sphingolipid-rich membrane microdomains termed "lipid rafts." However, the role of cholesterol and lipid rafts on chemokine binding and signaling through CCR5 remains unknown. We found that cholesterol extraction by hydroxypropyl-[beta]-cyclodextrin (BCD) significantly reduced the binding and signaling of macrophage inflammatory protein 1[beta] (MIP-1[beta]) using CCR5-expressing CEM-NKR T cells. Reloading treated cells with cholesterol but not 4-cholesten-3-one, an oxidized form of cholesterol, restored MIP-1[beta] binding to BCD-treated cells. Antibodies specific for distinct CCR5 epitopes lost their ability to bind to the cell surface after cholesterol extraction to varying degrees. Moreover, cells stained with fluorescently labeled MIP-1[beta] extensively colocalized with the GM1 lipid raft marker while using anti-CCR5 antibodies; most of CCR5 on these cells only partially colocalized with GM1, suggesting that active ligand binding facilitates receptor association with lipid rafts or that raft association promotes a higher affinity conformation of CCR5. Together, these data demonstrate that cholesterol and lipid rafts are important for the maintenance of the CCR5 conformation and are necessary for both the binding and function of this chemokine receptor. 10.1182/blood-2001-11-0087}, author = {Nguyen, Dzung H. and Taub, Dennis }, citeulike-article-id = {2602803}, doi = {http://dx.doi.org/10.1182/blood-2001-11-0087}, journal = {Blood}, keywords = {entry, hiv, raft, receptor}, month = {May}, number = {12}, pages = {4298--4306}, posted-at = {2008-03-27 17:45:22}, priority = {2}, title = {Cholesterol is essential for macrophage inflammatory protein 1beta binding and conformational integrity of CC chemokine receptor 5}, url = {http://dx.doi.org/10.1182/blood-2001-11-0087}, volume = {99}, year = {2002} }

TY - JOUR ID - citeulike:2602803 L3 - citeulike-article-id:2602803 TI - Cholesterol is essential for macrophage inflammatory protein 1beta binding and conformational integrity of CC chemokine receptor 5 JF - Blood VL - 99 IS - 12 SP - 4298 EP - 4306 N2 - The chemokine receptor, CCR5, is used as a human immunodeficiency virus coreceptor in combination with CD4 during transmission and early infection. CCR5 has been shown to be palmitoylated and targeted to cholesterol- and sphingolipid-rich membrane microdomains termed "lipid rafts." However, the role of cholesterol and lipid rafts on chemokine binding and signaling through CCR5 remains unknown. We found that cholesterol extraction by hydroxypropyl-[beta]-cyclodextrin (BCD) significantly reduced the binding and signaling of macrophage inflammatory protein 1[beta] (MIP-1[beta]) using CCR5-expressing CEM-NKR T cells. Reloading treated cells with cholesterol but not 4-cholesten-3-one, an oxidized form of cholesterol, restored MIP-1[beta] binding to BCD-treated cells. Antibodies specific for distinct CCR5 epitopes lost their ability to bind to the cell surface after cholesterol extraction to varying degrees. Moreover, cells stained with fluorescently labeled MIP-1[beta] extensively colocalized with the GM1 lipid raft marker while using anti-CCR5 antibodies; most of CCR5 on these cells only partially colocalized with GM1, suggesting that active ligand binding facilitates receptor association with lipid rafts or that raft association promotes a higher affinity conformation of CCR5. Together, these data demonstrate that cholesterol and lipid rafts are important for the maintenance of the CCR5 conformation and are necessary for both the binding and function of this chemokine receptor. 10.1182/blood-2001-11-0087 KW - entry KW - hiv KW - raft KW - receptor AU - Nguyen, Dzung AU - Taub, Dennis PY - 2002/05/29/ UR - http://dx.doi.org/10.1182/blood-2001-11-0087 ER -