Abstract
Endocytosis regulates the cell-surface expression of many seven transmembrane G-protein coupled receptors (7TM-GPCRs) and has been implicated in both desensitization following agonist-induced activation and resensitization (1,2). The internalization mechanism for many 7TM-GPCRs has not been clearly established. However, a clear paradigm is emerging for the β2-adrenergic receptor (β2AR) (1,2). For β2AR, agonist binding induces activation of heterotrimeric G proteins, which in turn activate one or more members of the family of G-protein coupled receptor kinases (GRKs). GRK-mediated phosphorylation of the receptor enhances the binding of β-arrestins, proteins that uncouple heterotrimeric G protein activation and link the receptor to clathrincoated pits by binding to clathrin heavy chains (1–3). Coated vesicles internalize the receptors and deliver them to endosomes, where they can be dephosphorylated and recycled to the cell surface (4).
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References
Ferguson S. S., Barak L. S., Zhang J., and Caron M. G. (1996) G-proteincoupled receptor regulation: role of G-protein-coupled receptor kinases and arrestins. Can. J. Physioi. Pharmacol. 74, 1095–1110.
Ferguson S. S., Downey III W. E., Colapietro A. M., Barak L. S., Menard L. and Caron M. G. (1996) Role of beta-arrestin in mediating agonist-promoted G protein-coupled receptor internalization. Science 271, 363–366.
Goodman 0. B. J., Krupnick J. G., Santinin F., Gurevich V. V., Penn R. B.. Gagnon A. W., Keen J. H., and Benovic J. L. (1996)β-Arrestin acts as aclathrin adaptor in endocytosis of the β2-adrenergic receptor. Nature 383, 447–450.
Krueger K. M., Daaka Y., Pitcher J. A., and Lefkowitz R. J. (1997) The role of sequestration in G protein-coupled receptor resensitization. Regulation of beta2-adrenergic receptor dephosphorylation by vesicular acidification. J. BioL Chem. 272, 5–8.
Amara A., Gall S. L., Schwartz O., Salamero J., Montes M., Loetscher P. Baggiolini M., Virelizier J. L., and Arenzana-Seisdedos F. (1997) HIV Coreceptor downregulation as antiviral principle: SDF-1 alpha-dependent internalization of the chemokine receptor CXCR4 contributes to inhibition of HIV replication. J. Exp. Med. 186, 139–146.
Signoret N., Oldridge J., Pelchen-Matthews A., Klasse P. J., Tran T., Brass L. F., Rosenkilde M. M., Schwartz T. W., Holmes W., Dallas W., Luther M. A., Wells T. N. C, Hoxie J. A., and Marsh M. (1997) Phorbol esters and SDF-1 induce rapid endocytosis and down modulation of the chemokine receptor CXCR4.J. Cell Biol 139, 651–664.
Solari R., Offord R. E., Remy S., Aubry J. P., Wells T. N. C, Whitehorn E., Oung T., and Proudfoot A. E. I. (1997) Receptor-mediated Endocytosis of CC chemokines. J. BioL Chem. 212, 9617–9620.
Samanta A. K., Oppenheim J. J., and Matsushima K. (1990) Interleukin 8 (monocyte-derived neutrophil chemotactic factor) dynamically regulates its own receptor expression on human neutrophils. J. BioL Chem. 265, 183–189.
Aramori I., Zhang J., Ferguson S. S., Bieniasz P. D., Cullen B. R., and Caron M. G. (1997) Molecular mechanism of desensitization of the chemokine receptor CCR-5: receptor signaling and internalization are dissociable from its role as an HIV-1 co-receptor. EMBO J. 16, 4606-1616.
Mack M., Luckow B., Nelson P. J., Cihak J., Simmons G., Clapham P. R., Signoret N., Marsh M., Borlat F., Stangassinger F., Wells T. N. C, Schlondorff D., and Proudfoot A. E. I. (1998) AOP-RANTES prevents CCR5 recycling: a novel inhibitory mechanism of HIV infectivity. J. Exp. Med. 187, 1215–1224.
Barak L. S., Ferguson S. S., Zhang J., Martenson C, Meyer T., and Caron M. G. (1997) Internal trafficking and surface mobility of a functionally intact beta2 adrenergic receptor-green fluorescent protein conjugate. Mol. Pharmacol. 51. 177–184.
Pelchen-Matthews A., Armes J. E., and Marsh M. (1989) Internalization and recycling of CD4 transfected into HeLa and NIH-3T3 cells. EMBO J. 8, 3641–3649.
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Signoret, N., Marsh, M. (2000). Analysis of Chemokine Receptor Endocytosis and Recycling. In: Proudfoot, A.E.I., Wells, T.N.C., Power, C.A. (eds) Chemokine Protocols. Methods in Molecular Biology, vol 138. Humana Press. https://doi.org/10.1385/1-59259-058-6:197
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DOI: https://doi.org/10.1385/1-59259-058-6:197
Publisher Name: Humana Press
Print ISBN: 978-0-89603-722-9
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