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Papers In Press, published online ahead of print January 18, 2001
West Australian Institute for Medical Research (WAIMR), Perth, W.A 6009
Corresponding Author: keidne{at}waimr.uwa.edu.au
The ability of G-protein coupled receptors (GPCRs) to interact to form new functional structures, either forming oligomers with themselves, or forming associations with other intracellular proteins, has important implications for the regulation of cellular events, however little is known about how this occurs. Here, we have employed a newly emerging technology, bioluminescence resonance energy transfer (BRET), used to study protein-protein interactions in living cells, to demonstrate that the thyrotropin releasing hormone receptor (TRHR) forms constitutive homo-oligomers. This formation of TRHR homo-oligomers in the absence of ligand, was shown by demonstration of an energy transfer between TRHR molecules fused to either donor, Renilla luciferase (Rluc) or acceptor, enhanced yellow fluorescent protein (EYFP) molecules. This interaction was shown to be specific, as energy transfer was not detected between co-expressed tagged TRHRs and either complementary tagged gonadotropin releasing hormone (GnRH) or b2-adrenergic receptors. Furthermore, generation of a BRET signal between the TRHRs could only be inhibited by co-expression of the wild-type TRHR and not by other GPCRs. Agonist-stimulation led to a time- and dose-dependent increase in the amount of energy transfer. Inhibition of receptor internalization by co-expression of dynamin mutant K44A did not affect the interaction between TRHRs, suggesting that clustering of receptors within clathrin-coated pits is not sufficient for energy transfer to occur. BRET also provided evidence for the agonist-induced oligomerization of another GPCR, the GnRH receptor (GnRHR), and the presence of an agonist-induced interaction of the adaptor protein, b-arrestin, with TRHR and the absence of an interaction of b-arrestin with GnRHR. This study supports the usefulness of BRET as a powerful tool for studying GPCR aggregations and receptor/protein interactions in general, and presents evidence that the functioning unit of TRH receptors exists as homomeric complexes.
J. Biol. Chem, 10.1074/jbc.M011311200
Submitted on December 15, 2000
Revised on January 16, 2001
Accepted on January 18, 2001
Constitutive and agonist-dependent homo-oligomerization of the thyrotropin-releasing hormone (TRH) receptor: detection in living cells using bioluminescence resonance energy transfer (BRET)
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|
|
|
|
|
|
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|
|
|
|
 |
|
 C. Blanpain, J.-M. Vanderwinden, J. Cihak, V. Wittamer, E. Le Poul, H. Issafras, M. Stangassinger, G. Vassart, S. Marullo, D. Schlondorff, et al. Multiple Active States and Oligomerization of CCR5 Revealed by Functional Properties of Monoclonal Antibodies Mol. Biol. Cell, February 1, 2002; 13(2): 723 - 737. [Abstract] [Full Text] [PDF]
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