Secretin Receptor Dimerization. Prototypic of Class B GPCR Behavior

Harikumar, Kaleeckal G.; Miller, Laurence J.

doi:10.1007/978-3-319-60174-8_10

Cited by 3 publications

(2 citation statements)

References 55 publications

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“…By means of dual color FRAP, they observed that the β 1 -adrenergic receptor (β 1 -AR) formed monomers that are unstable, in contrast to the β 2 -adrenergic receptor (β 2 -AR), which formed stable higher-order oligomers. The dimerization event in Class A GPCRs is more complicated than in other classes since there are multiple interfaces and owing to the fact that the interactions are transient in nature [56]. For D 2 R–D 2 R interactions, this was recently supported by modelling studies [23] and by studies indicating a dimer lifetime of only 68 ms [57,58].…”

Section: Discussionmentioning

confidence: 99%

Heterodimerization of Mu Opioid Receptor Protomer with Dopamine D2 Receptor Modulates Agonist-Induced Internalization of Mu Opioid Receptor

et al. 2019

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The interplay between the dopamine (DA) and opioid systems in the brain is known to modulate the additive effects of substances of abuse. On one hand, opioids serve mankind by their analgesic properties, which are mediated via the mu opioid receptor (MOR), a Class A G protein-coupled receptor (GPCR), but on the other hand, they pose a potential threat by causing undesired side effects such as tolerance and dependence, for which the exact molecular mechanism is still unknown. Using human embryonic kidney 293T (HEK 293T) and HeLa cells transfected with MOR and the dopamine D2 receptor (D2R), we demonstrate that these receptors heterodimerize, using an array of biochemical and biophysical techniques such as coimmunoprecipitation (co-IP), bioluminescence resonance energy transfer (BRET1), Fӧrster resonance energy transfer (FRET), and functional complementation of a split luciferase. Furthermore, live cell imaging revealed that D2LR, when coexpressed with MOR, slowed down internalization of MOR, following activation with the MOR agonist [D-Ala2, N-MePhe4, Gly-ol]-enkephalin (DAMGO).

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Section: Discussionmentioning

confidence: 99%

Heterodimerization of Mu Opioid Receptor Protomer with Dopamine D2 Receptor Modulates Agonist-Induced Internalization of Mu Opioid Receptor

et al. 2019

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“…Class B GPCRs have been shown to be intermediate between the class C GPCRs, exhibiting required and often covalent dimerization, to class A GPCRs, with such complexes being mostly transient [10]. Of the class B GPCRs, the human secretin receptor (hSecR) is prototypic, most extensively studied, and best characterized [13,15].…”

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confidence: 99%

Mechanistic insights from the atomic-level quaternary structure of short-lived GPCR oligomers in live cells

Raicu,

Stoneman,

Yokoi

et al. 2024

Preprint

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The functional significance of the interactions between proteins in living cells to form short-lived quaternary structures cannot be overemphasized. Yet, quaternary structure information is not captured by current methods, neither can those methods determine structure within living cells. The dynamic versatility, abundance, and functional diversity of G protein-coupled receptors (GPCRs) pose myriad challenges to existing technologies but also present these proteins as the ideal testbed for new technologies to investigate the complex inter-regulation of receptor-ligand, receptor-receptor, and receptor-downstream effector interfaces in living cells. Here, we present development and use of a novel method capable of overcoming existing challenges by combining distributions (or spectrograms) of FRET efficiencies from populations of fluorescently tagged proteins associating into oligomeric complexes in live cells with diffusion-like trajectories of FRET donors and acceptors obtained from molecular dynamics (MD) simulations. Our approach provides an atom-level picture of the binding interfaces within oligomers of the human secretin receptor (hSecR) in live cells and allows for extraction of mechanistic insights into the function of GPCRs oligomerization. This FRET-MD spectrometry approach is a robust platform for investigating protein-protein binding mechanisms and opens a new avenue for investigating stable as well as fleeting quaternary structures of any membrane proteins in living cells.

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