Detection of higher‐order G protein‐coupled receptor oligomers by a combined BRET–BiFC technique

Gandía, Jorge; Galino, Jorge; Amaral, Olavo B.; Soriano, Aroa; Lluı́s, Carme; Franco, Rafael; Ciruela, Francisco

doi:10.1016/j.febslet.2008.07.045

Cited by 90 publications

(70 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Each of them exhibited two intracellular domains that interacted in a specific manner with intracellular domains of the other two protomers through electrostatic interactions, leading to the assembly of dimers (A 2A -D 2 , A 2A -CB 1 , and CB 1 -D 2 ) but also to the formation of a A 2A -D 2 -CB 1 heterotrimer. Trimeric receptor complexes have been indeed identified (Gandia et al, 2008). They include the A 2A -D 2 -mGlu 5 (Cabello et al, 2009) heteroreceptor complex, the muscarinic M 2 homotrimer (Park and Wells, 2004), the dynamic Gal 1 -5-HT 1A -GPR 39 heterotrimer (Tena-Campos et al, 2015), and the putative Gal 1 -Gal 2 -5-HT 1A heterotrimer (Millón et al, 2016).…”

Section: Quaternary Structure Of Gpcr Complexesmentioning

confidence: 99%

“…Biological methods for identifying GPCR oligomers in cells and tissues or in recombinant mammalian expression systems presently include energy transfer-based methods [fluorescence resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET); Fernandez-Dueñas et al, 2012], bimolecular luminescence or fluorescence complementation (Gandia et al, 2008), total internal reflection fluorescence microscopy (Hern et al, 2010), fluorescence correlation spectroscopy (Chen et al, 2003), analysis of colocalization in immunohistochemical preparations (Agnati et al, 2005a), coimmunoprecipitation , assays based on bivalent ligands (Yekkirala et al, 2013), and in situ proximity ligation assays (Trifilieff et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

G protein-coupled receptor-receptor interactions give integrative dynamics to intercellular communication

Guidolin

Marcoli

Tortorella

et al. 2018

Reviews in the Neurosciences

View full text Add to dashboard Cite

Abstract:The proposal of receptor-receptor interactions (RRIs) in the early 1980s broadened the view on the role of G protein-coupled receptors (GPCR) in the dynamics of the intercellular communication. RRIs, indeed, allow GPCR to operate not only as monomers but also as receptor complexes, in which the integration of the incoming signals depends on the number, spatial arrangement, and order of activation of the protomers forming the complex. The main biochemical mechanisms controlling the functional interplay of GPCR in the receptor complexes are direct allosteric interactions between protomer domains. The formation of these macromolecular assemblies has several physiologic implications in terms of the modulation of the signaling pathways and interaction with other membrane proteins. It also impacts on the emerging field of connectomics, as it contributes to set and tune the synaptic strength. Furthermore, recent evidence suggests that the transfer of GPCR and GPCR complexes between cells via the exosome pathway could enable the target cells to recognize/decode transmitters and/or modulators for which they did not express the pertinent receptors. Thus, this process may also open the possibility of a new type of redeployment of neural circuits. The fundamental aspects of GPCR complex formation and function are the focus of the present review article.

show abstract

Section: Quaternary Structure Of Gpcr Complexesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

G protein-coupled receptor-receptor interactions give integrative dynamics to intercellular communication

Guidolin

Marcoli

Tortorella

et al. 2018

Reviews in the Neurosciences

View full text Add to dashboard Cite

show abstract

“…Due to its relative technical simplicity and the ability to use fluorescence microscopes for observation, a growing number of publications describe the use of BiFC to analyze protein-protein interactions. In addition to monitoring protein-protein interactions, this method has expanded to wider application, such as multicolor B iF C to investigate protein complexes (Hu & Kerppola, 2003;Kodama & Wada, 2009;Lee et al, 2008;Waadt et al, 2008), detection in vivo (Bracha-Drori et al, 2004;Walter et al, 2004) and combined with bioluminescence resonance energy transfer (BRET; Chen et al, 2008;Gandia et al, 2008;Xu et al, 2007). To date, several BiFC vectors dedicated to plant research have been constructed.…”

Section: Gateway Vectors For Bimolecular Fluorescence Complementationmentioning

confidence: 99%

Gateway Vectors for Plant Genetic Engineering: Overview of Plant Vectors, Application for Bimolecular Fluorescence Complementation (BiFC) and Multigene Construction

Tanaka¹,

Kimura²,

Hikino³

et al. 2012

Genetic Engineering - Basics, New Applications and Responsibilities

View full text Add to dashboard Cite

“…4), but the composition of the bands may be unclear, and the size under the conditions of electrophoresis may have little in common with that in the membrane. Larger oligomers also have been identified by approaches in which detection requires the colocalization of three or four proteins, each bearing a different tag (5)(6)(7)(8)(9)(10)(11), but such procedures place only a lower limit on the possible size of the array.…”

mentioning

confidence: 99%

Oligomeric Size of the M2 Muscarinic Receptor in Live Cells as Determined by Quantitative Fluorescence Resonance Energy Transfer

Pisterzi

Jansma

Georgiou

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Fluorescence resonance energy transfer (FRET), measured by fluorescence intensity-based microscopy and fluorescence lifetime imaging, has been used to estimate the size of oligomers formed by the M 2 muscarinic cholinergic receptor. The approach is based on the relationship between the apparent FRET efficiency within an oligomer of specified size (n) and the pairwise FRET efficiency between a single donor and a single acceptor (E). The M 2 receptor was fused at the N terminus to enhanced green or yellow fluorescent protein and expressed in Chinese hamster ovary cells. Emission spectra were analyzed by spectral deconvolution, and apparent efficiencies were estimated by donor-dequenching and acceptor-sensitized emission at different ratios of enhanced yellow fluorescent protein-M 2 receptor to enhanced green fluorescent protein-M 2 receptor. The data were interpreted in terms of a model that considers all combinations of donor and acceptor within a specified oligomer to obtain fitted values of E as follows: n ‫؍‬ 2, 0.495 ؎ 0.019; n ‫؍‬ 4, 0.202 ؎ 0.010; n ‫؍‬ 6, 0.128 ؎ 0.006; n ‫؍‬ 8, 0.093 ؎ 0.005. The pairwise FRET efficiency determined independently by fluorescence lifetime imaging was 0.20 -0.24, identifying the M 2 receptor as a tetramer. The strategy described here yields an explicit estimate of oligomeric size on the basis of fluorescence properties alone. Its broader application could resolve the general question of whether G protein-coupled receptors exist as dimers or larger oligomers. The size of an oligomer has functional implications, and such information can be expected to contribute to an understanding of the signaling process.Much evidence now indicates that G protein-coupled receptors can exist as oligomers (1, 2), a development that has implications for all aspects of GPCR 4 -mediated signaling. Among the many questions prompted by the emergence of such structures is that of oligomeric size. Although commonly referred to as dimers, oligomers of GPCRs have been detected most often by means of coimmunoprecipitation or resonance energy transfer (3). As typically applied, neither technique can distinguish dimers from larger oligomers. The latter have been identified on the basis of their electrophoretic mobility (reviewed in Ref. 4), but the composition of the bands may be unclear, and the size under the conditions of electrophoresis may have little in common with that in the membrane. Larger oligomers also have been identified by approaches in which detection requires the colocalization of three or four proteins, each bearing a different tag (5-11), but such procedures place only a lower limit on the possible size of the array.There have been comparatively few attempts to examine the oligomeric status of a GPCR in a more quantitative and explicit manner. Measurements of BRET at different ratios of acceptor to donor have pointed to dimers of the melatonin receptor (12), the ␤ 1 -and ␤ 2 -adrenergic receptors (13), the M 1 , M 2 , and M 3 muscarinic receptors (14), and the neurotensin receptor (15)....

show abstract

Detection of higher‐order G protein‐coupled receptor oligomers by a combined BRET–BiFC technique

Cited by 90 publications

References 22 publications

G protein-coupled receptor-receptor interactions give integrative dynamics to intercellular communication

G protein-coupled receptor-receptor interactions give integrative dynamics to intercellular communication

Gateway Vectors for Plant Genetic Engineering: Overview of Plant Vectors, Application for Bimolecular Fluorescence Complementation (BiFC) and Multigene Construction

Oligomeric Size of the M2 Muscarinic Receptor in Live Cells as Determined by Quantitative Fluorescence Resonance Energy Transfer

Contact Info

Product

Resources

About