scholarly journals Compartment-specific opioid receptor signaling is selectively modulated by different Dynorphin peptides

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Jennifer M Kunselman ◽  
Achla Gupta ◽  
Ivone gomes ◽  
Lakshmi A Devi ◽  
Manojkumar A Puthenveedu
Keyword(s):  
Opioid Receptor ◽  
Kappa Opioid Receptor ◽  
Neuroendocrine Cells ◽  
Striatal Neurons ◽  
Dynorphin A ◽  
Downstream Signaling ◽  
Intracellular Compartments ◽  
G Protein Coupled ◽  
Fine Tune ◽  
Signal Transduction Systems

Many signal transduction systems have an apparent redundancy built into them, where multiple physiological agonists activate the same receptors. Whether this is true redundancy, or whether this provides an as-yet unrecognized specificity in downstream signaling, is not well understood. We address this question using the kappa opioid receptor (KOR), a physiologically relevant G protein-coupled receptor (GPCR) that is activated by multiple members of the Dynorphin family of opioid peptides. We show that two related peptides, Dynorphin A and Dynorphin B, bind and activate KOR to similar extents in mammalian neuroendocrine cells and rat striatal neurons, but localize KOR to distinct intracellular compartments and drive different post-endocytic fates of the receptor. Strikingly, localization of KOR to the degradative pathway by Dynorphin A induces sustained KOR signaling from these compartments. Our results suggest that seemingly redundant endogenous peptides can fine-tune signaling by regulating the spatiotemporal profile of KOR signaling.

Compartment-specific opioid receptor signaling is selectively modulated by Dynorphin subtypes

2020 ◽  
Author(s):  
Jennifer M. Kunselman ◽  
Achla Gupta ◽  
Ivone Gomes ◽  
Lakshmi A. Devi ◽  
Manoj A. Puthenveedu
Keyword(s):  
Opioid Receptor ◽  
Opioid Peptides ◽  
Kappa Opioid Receptor ◽  
Endogenous Opioid Peptides ◽  
Endogenous Opioid ◽  
Spatial Profile ◽  
Downstream Signaling ◽  
G Protein Coupled ◽  
Fine Tune ◽  
Signal Transduction Systems

AbstractMany signal transduction systems have an apparent redundancy built into them, where multiple physiological agonists activate the same receptors. Whether this is true redundancy, or whether this provides as-yet unrecognized specificity in downstream signaling, is not well understood. We address this question using the kappa opioid receptor (KOR), a physiologically relevant G protein-coupled receptor (GPCR) that is activated by multiple members of the Dynorphin family of opioid peptides. We show that, although highly related Dynorphins bind and activate KOR to similar extents on the cell surface, they localize KOR to distinct subcellular compartments, dictate different post-endocytic fates of the receptor, and differentially induce KOR signaling from the degradative pathway. Our results show that seemingly redundant endogenous opioid peptides that are often co-released can in fact fine-tune signaling by differentially regulating the subcellular spatial profile of GPCR localization and signaling.

scholarly journals Molecular cloning of a rat κ opioid receptor reveals sequence similarities to the μ and δ opioid receptors

1993 ◽  
Vol 295 (3) ◽  
pp. 625-628 ◽  
Author(s):  
Y Chen ◽  
A Mestek ◽  
J Liu ◽  
L Yu
Keyword(s):  
Adenylate Cyclase ◽  
Opioid Receptor ◽  
Opioid Receptors ◽  
Kappa Opioid Receptor ◽  
Kappa Opioid ◽  
Brain Cdna Library ◽  
Inhibitory Coupling ◽  
Receptor Cdna ◽  
G Protein Coupled ◽  
Sequence Similarities

By screening a rat brain cDNA library using a cloned mu opioid receptor cDNA as probe, a clone was identified that is very similar to both the mu and delta opioid receptor sequences. Transient expression of this clone in COS-7 cells showed that it encodes a kappa opioid receptor, designated KOR-1, which is capable of high-affinity binding to kappa-selective ligands. Treatment of transfected cell membranes with bremazocine, a kappa-selective agonist, resulted in a 53% decrease in adenylate cyclase activity, indicating that this kappa opioid receptor displays inhibitory coupling to adenylate cyclase. Thus, one member from each of the three opioid receptor types, mu, kappa and delta, has been molecularly cloned. Analysis of sequence similarities among these opioid receptors, as well as between opioid receptors and other G-protein-coupled receptors, revealed regions of sequence conservation that may underlie the ligand-binding and functional specificities of opioid receptors.

scholarly journals Dual RXR motifs regulate nerve growth factor–mediated intracellular retention of the delta opioid receptor

2019 ◽  
Vol 30 (5) ◽  
pp. 680-690 ◽  
Author(s):  
Daniel J. Shiwarski ◽  
Stephanie E. Crilly ◽  
Andrew Dates ◽  
Manojkumar A. Puthenveedu
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Opioid Receptor ◽  
Mutational Analysis ◽  
Neuroendocrine Cells ◽  
Delta Opioid Receptor ◽  
Nerve Growth ◽  
Extracellular Signaling ◽  
Sequence Elements ◽  
Intracellular Compartments

The delta opioid receptor (DOR), a physiologically relevant prototype for G protein–coupled receptors, is retained in intracellular compartments in neuronal cells. This retention is mediated by a nerve growth factor (NGF)-regulated checkpoint that delays the export of DOR from the trans-Golgi network. How DOR is selectively retained in the Golgi, in the midst of dynamic membrane transport and cargo export, is a fundamental unanswered question. Here we address this by investigating sequence elements on DOR that regulate DOR surface delivery, focusing on the C-terminal tail of DOR that is sufficient for NGF-mediated regulation. By systematic mutational analysis, we define conserved dual bi-arginine (RXR) motifs that are required for NGF- and phosphoinositide-regulated DOR export from intracellular compartments in neuroendocrine cells. These motifs were required to bind the coatomer protein I (COPI) complex, a vesicle coat complex that mediates primarily retrograde cargo traffic in the Golgi. Our results suggest that interactions of DOR with COPI, via atypical COPI motifs on the C-terminal tail, retain DOR in the Golgi. These interactions could provide a point of regulation of DOR export and delivery by extracellular signaling pathways.

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

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Stephanie E Crilly ◽  
Wooree Ko ◽  
Zara Y Weinberg ◽  
Manojkumar A Puthenveedu
Keyword(s):  
G Protein ◽  
Opioid Receptor ◽  
Opioid Receptors ◽  
Neuronal Cells ◽  
Local Environment ◽  
Subcellular Location ◽  
Neuroendocrine Cells ◽  
Spatially Resolved ◽  
Δ Opioid Receptor ◽  
G Protein Coupled

The prevailing model for the variety in drug responses is that they stabilize distinct active states of their G protein-coupled receptor (GPCR) targets, allowing coupling to different effectors. However, whether the same ligand generates different GPCR active states based on the immediate environment of receptors is not known. Here we address this question using spatially resolved imaging of conformational biosensors that read out distinct active conformations of the δ-opioid receptor (DOR), a physiologically relevant GPCR localized to Golgi and the surface in neuronal cells. We show that Golgi and surface pools of DOR both inhibit cAMP, but engage distinct conformational biosensors in response to the same ligand in rat neuroendocrine cells. Further, DOR recruits arrestins on the surface but not the Golgi. Our results suggest that the local environment determines the active states of receptors for any given drug, allowing GPCRs to couple to different effectors at different subcellular locations.

14-β-Methyl-8-oxacyclorphan (BC-3016), a morphinan derivative with high affinity for kappa opioid receptor: comparison with dynorphin-A(1–13)

1986 ◽  
Vol 64 (6) ◽  
pp. 707-711 ◽  
Author(s):  
Simon Lemaire ◽  
Michel Dumont ◽  
François Jolicoeur ◽  
Bernard Belleau
Keyword(s):  
Guinea Pig ◽  
Rat Brain ◽  
Opioid Receptor ◽  
Opioid Receptors ◽  
Kappa Opioid Receptor ◽  
Dynorphin A ◽  
Mouse Vas Deferens ◽  
Structural Modifications ◽  
Potent Agonist ◽  
Mr 2266

14-β-Methyl-8-oxacyclorphan (BC-3016) was tested for its ability to depress the electrically evoked contractions of the guinea pig ileum (GPI) and of the mouse vas deferens (MVD) and to compete with the binding of prototype ligands selective for κ-, μ-, or δ-opioid receptors in membrane preparations of rat brain and guinea pig cerebellum. BC-3016 was a very potent agonist in the GPI and MVD preparations, with ID50 of 0.7 and 31 nM, respectively. The activity of levorphanol, a standard alkaloid related to BC-3016, was much lower in both assays with ID50 values of 44 and 86 nM, respectively. Conversely, the activity of BC-3016 was quite comparable to that of dynorphin-A(1–13) in both preparations. In the GPI assay, a putative κ-receptor antagonist, MR-2266, was 6.6 and 5.5 times more potent than naloxone in blocking the activity of BC-3016 and dynorphin-A(1–13), respectively. BC-3016 was also very potent in displacing bound [3H]ethylketocyclazocine ([3H]EKC) to membrane preparations of the guinea pig cerebellum, a brain component containing predominantly κ-opioid receptors (Ki of 0.58 nM). Its potency in the displacement of the bound μ-ligand, 3H-labelled (D-Ala2,MePhe4,Gly-OH5)-enkephalin ([3H]DAGO), to rat brain homogenates was somewhat lower (Ki of 0.8 nM) but still high when compared with its ability to displace the δ-ligand, 3H-labelled (D-Ser2, Thr6)-Leu-enkephalin ([3H]DSLET) to rat brain homogenates (Ki of 4.45 nM). The affinity of BC-3016 for the opioid receptor was 2.1-fold higher than that of U-50488H, a selective κ-opioid ligand. Finally, the relative potency of BC-3016 to bind each one of the receptor types was similar to that of dynorphin-A(1–13). These data indicate that the structural modifications of levorphanol leading to BC-3016 increases the affinity of the compound for the κ-opioid receptor and provides a compound whose profile of activity is similar in some respect to that of endogenous dynorphin-A(1–13).

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