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Molecular and Cellular Biology, July 2005, p. 5752-5762, Vol. 25, No. 13
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.13.5752-5762.2005

Ligand-Induced Partitioning of Human CXCR1 Chemokine Receptors with Lipid Raft Microenvironments Facilitates G-Protein-Dependent Signaling

Xuanmao Jiao,1 Ning Zhang,2 Xuehua Xu,1 Joost J. Oppenheim,2 and Tian Jin1*

Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, NIH, Rockville, Maryland 20852,1 Laboratory of Molecular Immunoregulation, National Cancer Institute, Frederick, Maryland 217012

Received 10 January 2005/ Returned for modification 2 February 2005/ Accepted 30 March 2005

Ligand binding to a chemokine receptor triggers signaling events through heterotrimeric G-proteins. The mechanisms underlying receptor-mediated G-protein activation in the heterogeneous microenvironments of the plasma membrane are unclear. Here, using live-cell fluorescence resonance energy transfer imaging to detect the proximity between CXCR1-cyan fluorescent protein (CFP) and fluorescence probes that label lipid raft or non-lipid raft microdomains and using fluorescence recovery after photobleaching analysis to measure the lateral diffusion of CXCR1-CFP, we found that interleukin-8 induces association between the receptors and lipid raft microenvironments. Disruption of lipid rafts impaired G-protein-dependent signaling, such as Ca2+ responses and phosphatidylinositol 3-kinase activation, but had no effect on ligand-binding function and did not completely abolish ligand-induced receptor phosphorylation. Our results suggest a novel mechanism by which ligand binding to CXCR1 promotes lipid raft partitioning of receptors and facilitates activation of heterotrimeric G-proteins.

* Corresponding author. Mailing address: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, NIH, 12441 Parklawn Drive, Twinbrook II, Rockville, MD 20852. Phone: (301) 480-1430. Fax: (301) 480-2618. E-mail: tjin{at}niaid.nih.gov.

Molecular and Cellular Biology, July 2005, p. 5752-5762, Vol. 25, No. 13
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.13.5752-5762.2005




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