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Ligand-dependent spatiotemporal signaling profiles of the mu-opioid receptor are controlled by distinct protein-protein interaction networks

ABSTRACT: Ligand-dependent differences in the regulation and internalization of the mu-opioid receptor (MOR) have been linked to the degree of severity of the side effects that limit the use of opiates in the treatment of pain. For example, activation of the MOR by morphine and DAMGO ([D-Ala2,N-MePhe4,Gly-ol]-enkephalin) causes distinct patterns of spatiotemporal signaling which are dependent on the distribution pattern of the receptor at the plasma membrane after activation. Morphine stimulation of MOR activates a Gβγ-protein kinase C (PKC)α-phosphorylation pathway that limits the distribution of the MOR and is associated with a sustained increase in cytosolic extracellular signal regulated kinase (ERK). In contrast, DAMGO causes a redistribution of the MOR at the plasma membrane (prior to receptor internalization), that facilitates transient activation of cytosolic and nuclear ERK. Here, we use proximity biotinylation proteomics to dissect the different protein-interaction networks that underlie the spatiotemporal signaling of morphine and DAMGO. We found that DAMGO, but not morphine, activates Rac1 (Ras‐related C3 botulinum toxin substrate 1). Rac1 activation is dependent on the scaffolding proteins IQ motif-containing GTPase-activating protein-1 (IQGAP1) and Crk-like protein (CRKL), and CRKL is required for the transient increase in nuclear ERK. In contrast, morphine increased the proximity of the MOR to desmosomes, specialized and highly-ordered membrane domains. Knockdown of desmosomal proteins junction plakoglobin (JUP) or desmocolin-1 (DSC1) switched the morphine spatiotemporal signaling profile to mimic that of DAMGO, revealing a transient increase in nuclear ERK. Identifying the MOR-interaction networks that control differential spatiotemporal signaling represents an important step towards understanding how signal compartmentalization contributes to the development of tolerance and dependence.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Kidney

SUBMITTER: Srgjan Chivchiristov

LAB HEAD: Michelle Louise Halls

PROVIDER: PXD013328 | Pride | 2019-09-26

REPOSITORIES: Pride

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Ligand-dependent spatiotemporal signaling profiles of the μ-opioid receptor are controlled by distinct protein-interaction networks.

Civciristov Srgjan S Huang Cheng C Liu Bonan B Marquez Elsa A EA Gondin Arisbel B AB Schittenhelm Ralf B RB Ellisdon Andrew M AM Canals Meritxell M Halls Michelle L ML

The Journal of biological chemistry 20190912 44

Ligand-dependent differences in the regulation and internalization of the μ-opioid receptor (MOR) have been linked to the severity of adverse effects that limit opiate use in pain management. MOR activation by morphine or [d-Ala2,N-MePhe4, Gly-ol]enkephalin (DAMGO) causes differences in spatiotemporal signaling dependent on MOR distribution at the plasma membrane. Morphine stimulation of MOR activates a Gαi/o-Gβγ-protein kinase C (PKC) α phosphorylation pa ...[more]

PMID: 31515267

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