Conformational specificity of opioid receptors is determined by subcellular location irrespective of agonist

Crilly, Stephanie E; Ko, Wooree; Weinberg, Zara Y.; Puthenveedu, Manojkumar A.

doi:10.7554/elife.67478

Cited by 27 publications

(21 citation statements)

References 80 publications

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“…In particular, the efficiency of Gs coupling was markedly augmented by either high cholesterol or absence of the primary (Gq/11) transducers, demonstrating that the local environment of the receptor within a cell is also critical for the mode of transducer engagement. Differential transducer engagement has been reported for different cellular compartments, linked to different receptor conformations [ 46 ], and our current observations expand the potential mechanisms for such observations. Our work supports a model for G protein interaction where the extent of GPCR conformational change that occurs upon agonist activation is intrinsic to the individual receptor, but where conformational dynamics within the primary activated state are likely to play a substantial role in G protein coupling and activation.…”

Section: Discussionsupporting

confidence: 83%

Structures of the human cholecystokinin 1 (CCK1) receptor bound to Gs and Gq mimetic proteins provide insight into mechanisms of G protein selectivity

et al. 2021

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G protein–coupled receptors (GPCRs) are critical regulators of cellular function acting via heterotrimeric G proteins as their primary transducers with individual GPCRs capable of pleiotropic coupling to multiple G proteins. Structural features governing G protein selectivity and promiscuity are currently unclear. Here, we used cryo-electron microscopy (cryo-EM) to determine structures of the cholecystokinin (CCK) type 1 receptor (CCK1R) bound to the CCK peptide agonist, CCK-8 and 2 distinct transducer proteins, its primary transducer Gq, and the more weakly coupled Gs. As seen with other Gq/11–GPCR complexes, the Gq–α5 helix (αH5) bound to a relatively narrow pocket in the CCK1R core. Surprisingly, the backbone of the CCK1R and volume of the G protein binding pocket were essentially equivalent when Gs was bound, with the Gs αH5 displaying a conformation that arises from “unwinding” of the far carboxyl-terminal residues, compared to canonically Gs coupled receptors. Thus, integrated changes in the conformations of both the receptor and G protein are likely to play critical roles in the promiscuous coupling of individual GPCRs.

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Section: Discussionsupporting

confidence: 83%

Structures of the human cholecystokinin 1 (CCK1) receptor bound to Gs and Gq mimetic proteins provide insight into mechanisms of G protein selectivity

et al. 2021

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“…The present results, together with previous studies, establish that ORs in the Golgi apparatus are signaling competent as they engage transducer proteins (G proteins, GRKs) and inhibit cAMP production in response to permeant agonists ( 7, 32 ). Yet the transcriptomic and phosphoproteomic analyses delineate prominent differences in the downstream effects of Golgi-localized DOR relative to the PM receptor pool.…”

Section: Discussionsupporting

confidence: 85%

“…The location-dependent binding of β-arrestin to active ORs was reminiscent of the recruitment pattern of a specific mG probe, termed mGsi, which had recently been shown to bind to active DOR in the PM, but not in the Golgi apparatus ( 32 ). mGsi consists of the Ras-like domain of Gαs, but contains selectivity determining residues of Gαi in the C-terminal α5-helix (Suppl.…”

Section: Resultsmentioning

confidence: 99%

“…We here discover that Golgi-localized MOR and DOR are phosphorylated by GRK2/3 in an agonist-dependent manner, however they do not drive β-arrestin recruitment, contrasting with PM ORs in the same cell. It conveys that additional, location-specific determinants regulate β-arrestin–OR binding ( 32 ). β-arrestins can directly interact with membrane lipids via loops in the C-domain, which contributes to the stability of select β-arrestin–GPCR complexes ( 53, 54 ).…”

Section: Discussionmentioning

confidence: 99%

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Subcellular location defines GPCR signal transduction

Radoux-Mergault

Oberhauser

Aureli

et al. 2022

Preprint

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G protein-coupled receptors in intracellular organelles can be activated in response to membrane permeant ligands, which contributes to the diversity and specificity of agonist action. The opioid receptors (ORs) provide a striking example, where opioid drugs activate ORs in the Golgi apparatus within seconds of drug addition. Till date, our knowledge on the signaling of intracellular GPCRs remains incomplete and it is unknown if the downstream effects triggered by ORs in plasma membrane and Golgi apparatus differ. To address this gap, we first assess the recruitment of signal transducers to ORs in both compartments. We find that Golgi-localized ORs couple to Gai/o probes and are phosphorylated by GPCR kinases (GRK2/3), but unlike plasma membrane receptors, do not recruit b-arrestin or a specific Ga probe. Subsequent molecular dynamics simulations with OR-transducer complexes in model bilayers mimicking plasma membrane or Golgi composition reveal that the lipid environment promotes location selective coupling. Unbiased global analyses then show that OR activation in the plasma membrane and Golgi apparatus has strikingly different downstream effects on transcription and protein phosphorylation. Taken together, the study delineates OR signal transduction with unprecedented spatial resolution and reveals that the subcellular location defines the signaling effect of opioid drugs.

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“…Endocytosis therefore stringently restricts the number of plasma membrane receptors available for signaling. Interestingly, there have been several reports suggesting that the internalized receptors also mediate signaling and the signaling mediated by the internalized GPCR could be different from that on the plasma membrane ( Irannejad et al, 2013 ; Irannejad et al, 2015 ; Lohse and Hofmann, 2015 ; Bowman et al, 2016 ; Eichel and von Zastrow, 2018 ; Jong et al, 2018 ; Thomsen et al, 2018 ; Lobingier and von Zastrow, 2019 ; Crilly et al, 2021 ; Crilly and Puthenveedu, 2021 ; Kumar and Puthenveedu, 2022 ).…”

Section: Endocytosis and Cellular Trafficking: New Paradigm In Slos R...mentioning

confidence: 99%

Smith-Lemli-Opitz syndrome: A pathophysiological manifestation of the Bloch hypothesis

Chattopadhyay

Sharma²

2023

Front. Mol. Biosci.

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The biosynthesis of cholesterol, an essential component of higher eukaryotic membranes, was worked out by Konrad Bloch (and Feodor Lynen) in the 1960s and they received the Nobel Prize around that time in recognition of their pioneering contributions. An elegant consequence of this was a hypothesis proposed by Konrad Bloch (the Bloch hypothesis) which suggests that each subsequent intermediate in the cholesterol biosynthesis pathway is superior in supporting membrane function in higher eukaryotes relative to its precursor. In this review, we discuss an autosomal recessive metabolic disorder, known as Smith-Lemli-Opitz syndrome (SLOS), associated with a defect in the Kandutsch-Russell pathway of cholesterol biosynthesis that results in accumulation of the immediate precursor of cholesterol in its biosynthetic pathway (7-dehydrocholesterol) and an altered cholesterol to total sterol ratio. Patients suffering from SLOS have several developmental, behavioral and cognitive abnormalities for which no drug is available yet. We characterize SLOS as a manifestation of the Bloch hypothesis and review its molecular etiology and current treatment. We further discuss defective Hedgehog signaling in SLOS and focus on the role of the serotonin1A receptor, a representative neurotransmitter receptor belonging to the GPCR family, in SLOS. Notably, ligand binding activity and cellular signaling of serotonin1A receptors are impaired in SLOS-like condition. Importantly, cellular localization and intracellular trafficking of the serotonin1A receptor (which constitute an important determinant of a GPCR cellular function) are compromised in SLOS. We highlight some of the recent developments and emerging concepts in SLOS pathobiology and suggest that novel therapies based on trafficking defects of target receptors could provide new insight into treatment of SLOS.

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Conformational specificity of opioid receptors is determined by subcellular location irrespective of agonist

Cited by 27 publications

References 80 publications

Structures of the human cholecystokinin 1 (CCK1) receptor bound to Gs and Gq mimetic proteins provide insight into mechanisms of G protein selectivity

Structures of the human cholecystokinin 1 (CCK1) receptor bound to Gs and Gq mimetic proteins provide insight into mechanisms of G protein selectivity

Subcellular location defines GPCR signal transduction

Smith-Lemli-Opitz syndrome: A pathophysiological manifestation of the Bloch hypothesis

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