Probing novel GPCR interactions using a combination of FRET and TIRF
- PMID: 20798821
- PMCID: PMC2928313
- DOI: 10.4161/cib.3.4.11764
Probing novel GPCR interactions using a combination of FRET and TIRF
Abstract
Recent work on G-protein coupled receptors (GPCRs) has highlighted the importance of homo- and heterodimerization in all areas of GPCR function, including trafficking, signaling and desensitization. Novel GPCR dimers and even high-order oligomers are constantly being discovered. Advances in techniques such as fluorescent microscopy have improved our ability to detect these interactions. As GPCRs represent the largest class of transmembrane signaling molecules in biology, these new insights into their function could vastly improve our understanding of the complex physiological role GPCRs play in cellular signaling. Utilizing a combination of classic biochemical approaches and newer techniques such as fluorescence resonance energy transfer (FRET) and total internal reflection fluorescence microscopy (TIRF), we recently demonstrated a novel interaction between M(2) muscarinic receptors and GABA(B) receptors. In this addendum, we address technical aspects of combining FRET and TIRF to study GPCR interactions and further discuss the physiological implications of the M(2)-GABA(B) heterodimer.
Keywords: FRET; GABAB; GPCR; TIRF; cAMP; dimerization; potassium channel.
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Comment on
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Direct interaction of GABAB receptors with M2 muscarinic receptors enhances muscarinic signaling.J Neurosci. 2009 Dec 16;29(50):15796-809. doi: 10.1523/JNEUROSCI.4103-09.2009. J Neurosci. 2009. PMID: 20016095 Free PMC article.
References
-
- Margeta-Mitrovic M, Jan YN, Jan LY. A trafficking checkpoint controls GABA(B) receptor heterodimerization. Neuron. 2000;27:97–106. - PubMed
-
- Chang W, Tu C, Cheng Z, Rodriguez L, Chen TH, Gassmann M, et al. Complex formation with the Type Bgamma-aminobutyric acid receptor affects the expression and signal transduction of the extracellular calcium-sensing receptor. Studies with HEK-293 cells and neurons. J Biol Chem. 2007;282:25030–25040. - PubMed
-
- van Koppen CJ, Kaiser B. Regulation of muscarinic acetylcholine receptor signaling. Pharmacol Ther. 2003;98:197–220. - PubMed
-
- Wang D, Sun X, Bohn LM, Sadee W. Opioid receptor homo- and heterodimerization in living cells by quantitative bioluminescence resonance energy transfer. Mol Pharmacol. 2005;67:2173–2184. - PubMed
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