HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 37, 35354-35361, September 12, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/37/35354    most recent M305607200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Floyd, D. H. Articles by Baranski, T. J. Search for Related Content PubMed PubMed Citation Articles by Floyd, D. H. Articles by Baranski, T. J. Social Bookmarking                      What's this?

C5a Receptor Oligomerization

II. FLUORESCENCE RESONANCE ENERGY TRANSFER STUDIES OF A HUMAN G PROTEIN-COUPLED RECEPTOR EXPRESSED IN YEAST^*

Desiree H. Floyd , Adi Geva , Stephen P. Bruinsma , Mark C. Overton , Kendall J. Blumer  and Thomas J. Baranski  ¶

From the Departments of Medicine, Molecular Biology, and Pharmacology and the Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110

Recent studies demonstrate that members of the superfamily of G protein-coupled receptors (GPCRs) form oligomers both in vitro and in vivo. The mechanisms by which GPCRs oligomerize and the roles of accessory proteins in this process are not well understood. We used disulfide-trapping experiments to show that C5a receptors, expressed in mammalian cells, reside in membranes as oligomers (Klco, J. M., Lassere, T. B., and Baranski, T. J. (2003) J. Biol. Chem. 278, 35345–35353). To begin to address how C5a receptors form oligomers, we now use fluorescence resonance energy transfer experiments on human C5a receptors expressed in the lower eukaryote Saccharomyces cerevisiae. C5a receptors tagged with variants of the green fluorescent protein display energy transfer in intact yeast, demonstrating that mammalian accessory proteins are not required for C5a receptor oligomerization. In both intact yeast cells and membrane preparations, agonist does not affect FRET efficiency, and little energy transfer is observed between the C5a receptor and a co-expressed yeast pheromone receptor (encoded by STE2), indicating that C5a receptor oligomerization is both receptor-specific and constitutive. FRET studies performed on fractionated membranes demonstrate similar levels of energy transfer between tagged C5a receptors in endoplasmic reticulum compared with plasma membrane, and urea washing of membranes has little effect on the extent of energy transfer. The oligomerization of C5a receptors expressed in yeast displays characteristics similar to those observed for other GPCRs studied in mammalian cells. This model system should prove useful for further studies to define mechanisms of oligomerization of mammalian GPCRs.

Received for publication, May 28, 2003

^* The research was supported in part by grants from the Edward J. Mallinckrodt, Jr. Foundation (to T. J. B.), American Cancer Society Grant IRG-58-010-43 (to T. J. B.), the Culpeper Award, the Rockefeller Brothers Fund (to T. J. B.), National Institutes of Health Grant GM63720-01 (to T. J. B.), and a PhRMA predoctoral fellowship award (to S. P. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: Depts. of Medicine, Molecular Biology, and Pharmacology, Washington University School of Medicine, Box 8103, 660 S. Euclid Ave., St. Louis, MO 63110. Tel.: 314-747-3997; Fax: 314-362-7058; E-mail: baranski{at}pcg.wustl.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M.-J. Rabiet, E. Huet, and F. Boulay Complement Component 5a Receptor Oligomerization and Homologous Receptor Down-regulation J. Biol. Chem., November 7, 2008; 283(45): 31038 - 31046. [Abstract] [Full Text] [PDF]

				L. Szidonya, M. Cserzo, and L. Hunyady Dimerization and oligomerization of G-protein-coupled receptors: debated structures with established and emerging functions J. Endocrinol., March 1, 2008; 196(3): 435 - 453. [Abstract] [Full Text] [PDF]

				I. Hughes, M. Saito, P. H. Schlesinger, and D. M. Ornitz Otopetrin 1 activation by purinergic nucleotides regulates intracellular calcium PNAS, July 17, 2007; 104(29): 12023 - 12028. [Abstract] [Full Text] [PDF]

				M. L. Matsumoto, K. Narzinski, P. D. Kiser, G. V. Nikiforovich, and T. J. Baranski A Comprehensive Structure-Function Map of the Intracellular Surface of the Human C5a Receptor: I. IDENTIFICATION OF CRITICAL RESIDUES J. Biol. Chem., February 2, 2007; 282(5): 3105 - 3121. [Abstract] [Full Text] [PDF]

				P. Savi, J.-L. Zachayus, N. Delesque-Touchard, C. Labouret, C. Herve, M.-F. Uzabiaga, J.-M. Pereillo, J.-M. Culouscou, F. Bono, P. Ferrara, et al. The active metabolite of Clopidogrel disrupts P2Y12 receptor oligomers and partitions them out of lipid rafts PNAS, July 18, 2006; 103(29): 11069 - 11074. [Abstract] [Full Text] [PDF]

				M. M. C. Kong, T. Fan, G. Varghese, B. F. O'Dowd, and S. R. George Agonist-Induced Cell Surface Trafficking of an Intracellularly Sequestered D1 Dopamine Receptor Homo-Oligomer Mol. Pharmacol., July 1, 2006; 70(1): 78 - 89. [Abstract] [Full Text] [PDF]

				A. Singh, S. Severance, N. Kaur, W. Wiltsie, and D. J. Kosman Assembly, Activation, and Trafficking of the Fet3p{middle dot}Ftr1p High Affinity Iron Permease Complex in Saccharomyces cerevisiae J. Biol. Chem., May 12, 2006; 281(19): 13355 - 13364. [Abstract] [Full Text] [PDF]

				M. C. Overton, S. L. Chinault, and K. J. Blumer Oligomerization of G-Protein-Coupled Receptors: Lessons from the Yeast Saccharomyces cerevisiae Eukaryot. Cell, December 1, 2005; 4(12): 1963 - 1970. [Full Text] [PDF]

				D. Calebiro, T. de Filippis, S. Lucchi, C. Covino, S. Panigone, P. Beck-Peccoz, D. Dunlap, and L. Persani Intracellular entrapment of wild-type TSH receptor by oligomerization with mutants linked to dominant TSH resistance Hum. Mol. Genet., October 15, 2005; 14(20): 2991 - 3002. [Abstract] [Full Text] [PDF]

				E. M. Pietila, J. T. Tuusa, P. M. Apaja, J. T. Aatsinki, A. E. Hakalahti, H. J. Rajaniemi, and U. E. Petaja-Repo Inefficient Maturation of the Rat Luteinizing Hormone Receptor: A PUTATIVE WAY TO REGULATE RECEPTOR NUMBERS AT THE CELL SURFACE J. Biol. Chem., July 15, 2005; 280(28): 26622 - 26629. [Abstract] [Full Text] [PDF]

				G. Milligan G Protein-Coupled Receptor Dimerization: Function and Ligand Pharmacology Mol. Pharmacol., July 1, 2004; 66(1): 1 - 7. [Abstract] [Full Text] [PDF]

				J. L. Hansen, J. Theilade, S. Haunso, and S. P. Sheikh Oligomerization of Wild Type and Nonfunctional Mutant Angiotensin II Type I Receptors Inhibits G{alpha}q Protein Signaling but Not ERK Activation J. Biol. Chem., June 4, 2004; 279(23): 24108 - 24115. [Abstract] [Full Text] [PDF]

				M. C. Overton, S. L. Chinault, and K. J. Blumer Oligomerization, Biogenesis, and Signaling Is Promoted by a Glycophorin A-like Dimerization Motif in Transmembrane Domain 1 of a Yeast G Protein-coupled Receptor J. Biol. Chem., December 5, 2003; 278(49): 49369 - 49377. [Abstract] [Full Text] [PDF]

				J. M. Klco, T. B. Lassere, and T. J. Baranski C5a Receptor Oligomerization: I. DISULFIDE TRAPPING REVEALS OLIGOMERS AND POTENTIAL CONTACT SURFACES IN A G PROTEIN-COUPLED RECEPTOR J. Biol. Chem., September 12, 2003; 278(37): 35345 - 35353. [Abstract] [Full Text] [PDF]