Oligomeric potential of the M<sub>2</sub> muscarinic cholinergic receptor

Park, Paul S.‐H.; Wells, James W.

doi:10.1111/j.1471-4159.2004.02536.x

Cited by 55 publications

(46 citation statements)

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“…The data indicate that the receptor exists as a tetramer, as suggested previously on the basis of electrophoretic mobility (4,5,18), copurification (4,5,18), and noncompetitive effects in the binding of agonists and antagonists (4, 18 -20). Similar studies on other GPCRs can be expected to reveal the degree of oligomeric diversity within the rhodopsin-like subgroup and across the family at large.…”

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confidence: 51%

“…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.…”

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confidence: 99%

“…In each case, however, the data were analyzed in terms of a model developed for gramicidin and based on the notion of a dynamic equilibrium between the monomeric and oligomeric states (16). Measurements of the efficiency at which differently tagged adducts were coimmunoprecipitated from extracts of coinfected Sf9 cells suggested that the M 2 muscarinic receptor is at least a trimer (5), but the value depended upon properties that were difficult to measure or control.…”

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confidence: 99%

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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

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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)....

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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

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“…By combining BiFC and BiLC with BRET measurements, we now showed that heteromeric complexes between BILF1 and CXCR4 consist of at least four interacting GPCR proteins at the same time. Such oligomeric organization has been observed for native rhodopsin in disc membranes of rod outer segments by using atomic force microscopy (11) but have also been observed between class C GPCRs GABA B1 and GABA B2 (36) and between various class A GPCRs using various modifications and/or combinations of resonance energy transfer-based methods as well as cross-linking experiments (22,(37)(38)(39)(40)(41). Hitherto, however, the functional significance and/or necessity of oligomeric receptor complexes versus receptor dimers have remained puzzling.…”

Section: Discussionmentioning

confidence: 99%

The Epstein-Barr Virus-encoded G Protein-coupled Receptor BILF1 Hetero-oligomerizes with Human CXCR4, Scavenges Gαi Proteins, and Constitutively Impairs CXCR4 Functioning

Nijmeijer¹,

Leurs²,

Smit³

et al. 2010

Journal of Biological Chemistry

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Cells express distinct G protein-coupled receptor (GPCR)subtypes on their surface, allowing them to react to a corresponding variety of extracellular stimuli. Cross-regulation between different ligand-GPCR pairs is essential to generate appropriate physiological responses. GPCRs can physically affect each other's functioning by forming heteromeric complexes, whereas cross-regulation between activated GPCRs also occurs through integration of shared intracellular signaling networks. Human herpesviruses utilize virally encoded GPCRs to hijack cellular signaling networks for their own benefit. Previously, we demonstrated that the Epstein-Barr virus-encoded GPCR BILF1 forms heterodimeric complexes with human chemokine receptors. Using a combination of bimolecular complementation and bioluminescence resonance energy transfer approaches, we now show the formation of hetero-oligomeric complexes between this viral GPCR and human CXCR4. BILF1 impaired CXCL12 binding to CXCR4 and, consequently, also CXCL12-induced signaling. In contrast, the G protein uncoupled mutant BILF1-K 3.50 A affected CXCL12-induced CXCR4 signaling to a much lesser extent, indicating that BILF1-mediated CXCR4 inhibition is a consequence of its constitutive activity. Co-expression of G␣ i1 with BILF1 and CXCR4 restored CXCL12-induced signaling. Likewise, BILF1 formed heteromers with the human histamine H 4 receptor (H 4 R). BILF1 inhibited histamine-induced G␣ i -mediated signaling by H 4 R, however, without affecting histamine binding to this receptor. These data indicate that functional cross-regulation of G␣ i -coupled GPCRs by BILF1 is at the level of G proteins, even though these GPCRs are assembled in hetero-oligomeric complexes.

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“…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). Tetrameric assemblies of β 2 -adrenergic receptors were demonstrated by the group of Kobilka to occur spontaneously following reconstitution into phospholipid vesicles (Fung et al, 2009), suggesting that β 2 -adrenergic receptor oligomerization is an intrinsic property of the receptor.…”

Section: Quaternary Structure Of Gpcr Complexesmentioning

confidence: 99%

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

Guidolin

Marcoli

Tortorella

et al. 2018

Reviews in the Neurosciences

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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.

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Oligomeric potential of the M₂ muscarinic cholinergic receptor

Cited by 55 publications

References 47 publications

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

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

The Epstein-Barr Virus-encoded G Protein-coupled Receptor BILF1 Hetero-oligomerizes with Human CXCR4, Scavenges Gαi Proteins, and Constitutively Impairs CXCR4 Functioning

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

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Oligomeric potential of the M2 muscarinic cholinergic receptor

Cited by 55 publications

References 47 publications

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

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

The Epstein-Barr Virus-encoded G Protein-coupled Receptor BILF1 Hetero-oligomerizes with Human CXCR4, Scavenges Gαi Proteins, and Constitutively Impairs CXCR4 Functioning

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

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Oligomeric potential of the M₂ muscarinic cholinergic receptor