How the ubiquitous GPCR receptor family selectively activates signalling pathways

Capper, Michael J.; Wacker, Daniel

doi:10.1038/d41586-018-05503-4

Cited by 29 publications

(17 citation statements)

References 14 publications

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“…Recent cryo-electron microscopy (cryoEM) studies reporting the structures of G i/o bound GPCRs, such as μ-opioid 75 , adenosine A 1 76 , 5HT 1B 77 , and light receptor rhodopsin 78 , determine the interaction of these receptors with G i or G o and suggest the conformational re-arrangements on the GPCR cytoplasmic site may affect the binding of specific G proteins. Interestingly, they found different interactions of G i/o bound GPCRs and Gβ subunits 79 . However, the role of Gβγ in GPCRs-G protein specificity is unclear in these studies due to the modification of the proteins and the resolution of cryoEM structures.…”

Section: Discussionmentioning

confidence: 99%

The in vivo specificity of synaptic Gβ and Gγ subunits to the α2a adrenergic receptor at CNS synapses

Yim

Betke

Gilsbach

et al. 2019

Sci Rep

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G proteins are major transducers of signals from G-protein coupled receptors (GPCRs). They are made up of α, β, and γ subunits, with 16 Gα, 5 Gβ and 12 Gγ subunits. Though much is known about the specificity of Gα subunits, the specificity of Gβγs activated by a given GPCR and that activate each effector in vivo is not known. Here, we examined the in vivo Gβγ specificity of presynaptic α2a-adrenergic receptors (α2aARs) in both adrenergic (auto-α2aARs) and non-adrenergic neurons (hetero-α2aARs) for the first time. With a quantitative MRM proteomic analysis of neuronal Gβ and Gγ subunits, and co-immunoprecipitation of tagged α2aARs from mouse models including transgenic FLAG-α2aARs and knock-in HA-α2aARs, we investigated the in vivo specificity of Gβ and Gγ subunits to auto-α2aARs and hetero-α2aARs activated with epinephrine to understand the role of Gβγ specificity in diverse physiological functions such as anesthetic sparing, and working memory enhancement. We detected Gβ2, Gγ2, Gγ3, and Gγ4 with activated auto α2aARs, whereas we found Gβ4 and Gγ12 preferentially interacted with activated hetero-α2aARs. Further understanding of in vivo Gβγ specificity to various GPCRs offers new insights into the multiplicity of genes for Gβ and Gγ, and the mechanisms underlying GPCR signaling through Gβγ subunits.

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Section: Discussionmentioning

confidence: 99%

The in vivo specificity of synaptic Gβ and Gγ subunits to the α2a adrenergic receptor at CNS synapses

Yim

Betke

Gilsbach

et al. 2019

Sci Rep

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“…This smaller TM6 displacement has been suggested to be a feature of GPCR-G i/o complexes compared to the larger TM6 displacement required to accommodate the bulkier α5 helix of Gα s (Extended Data Fig. 7a) 24 .…”

Section: Structures Of Hntsr1 In C and Nc Statesmentioning

confidence: 92%

Conformational transitions of a neurotensin receptor 1–Gi1 complex

Kato

Zhang

et al. 2019

Nature

214

250

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The neurotensin receptor 1 (NTSR1) is a G-protein-coupled receptor (GPCR) that engages multiple G-protein subtypes and is involved in regulation of blood pressure, body temperature, weight, and response to pain. Here we present 3-Å structures of the human NTSR1 in complex with the agonist JMV449 and the heterotrimeric G i1 protein in two conformations (C state and NC Reprints and permissions information is available at www.nature.com/reprints.Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

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“…G protein-coupled receptors (GPCRs) play a central role in numerous signal transduction pathways across the cell membrane, which are initiated by binding of extracellular ligands to the receptor and subsequent activation of different intracellular signaling pathways. Due to this central role in various biochemical signal transduction cascades, GPCRs are of high pharmacological relevance [ 1 ].…”

Section: Introductionmentioning

confidence: 99%

Probing the Y2 Receptor on Transmembrane, Intra- and Extra-Cellular Sites for EPR Measurements

et al. 2020

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The function of G protein-coupled receptors is intrinsically linked to their conformational dynamics. In conjugation with site-directed spin labeling, electron paramagnetic resonance (EPR) spectroscopy provides powerful tools to study the highly dynamic conformational states of these proteins. Here, we explored positions for nitroxide spin labeling coupled to single cysteines, introduced at transmembrane, intra- and extra-cellular sites of the human neuropeptide Y2 receptor. Receptor mutants were functionally analyzed in cell culture system, expressed in Escherichia coli fermentation with yields of up to 10 mg of purified protein per liter expression medium and functionally reconstituted into a lipid bicelle environment. Successful spin labeling was confirmed by a fluorescence assay and continuous wave EPR measurements. EPR spectra revealed mobile and immobile populations, indicating multiple dynamic conformational states of the receptor. We found that the singly mutated positions by MTSL ((1-oxyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrol-3-yl) methyl methanesulfonothioate) have a water exposed immobilized conformation as their main conformation, while in case of the IDSL (bis(1-oxyl-2,2,5,5-tetramethyl-3-imidazolin-4-yl) disulfide) labeled positions, the main conformation are mainly of hydrophobic nature. Further, double cysteine mutants were generated and examined for potential applications of distance measurements by double electron–electron resonance (DEER) pulsed EPR technique on the receptor.

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How the ubiquitous GPCR receptor family selectively activates signalling pathways

Cited by 29 publications

References 14 publications

The in vivo specificity of synaptic Gβ and Gγ subunits to the α2a adrenergic receptor at CNS synapses

The in vivo specificity of synaptic Gβ and Gγ subunits to the α2a adrenergic receptor at CNS synapses

Conformational transitions of a neurotensin receptor 1–Gi1 complex

Probing the Y2 Receptor on Transmembrane, Intra- and Extra-Cellular Sites for EPR Measurements

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