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Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis
Published on: July 3, 2015
Ubiquitination of the μ-opioid receptor regulates receptor internalization without affecting Gi/o-mediated
Kentaro Miyoshi1, Satoshi Shimizu2, Atsuko Shiraki1
1Department of Anesthesia, Kyoto University Hospital, 54 Shogoin-kawahara-cho, Sakyo-ku, Kyoto City, Kyoto Prefecture, Japan.
Abstract:
Opioids are highly potent analgesics but develop tolerance. Previous studies have focused on phosphorylation of the μ-opioid receptor as it is involved in maintaining cellular sensitivity via desensitization, recycling, and degradation of the activated receptor. Recently, ubiquitination, another form of posttranslational modification has attracted attention in terms of triggering intracellular signaling and regulation of the activated receptor. Here, we generated a ubiquitination-deficient mutant of the μ-opioid receptor to investigate whether ubiquitination is involved in driving Gi/o-mediated analgesic signaling, receptor desensitization or subsequent receptor internalization. Our study shows that the Gi/o pathway and receptor phosphorylation do not require ubiquitination. Instead, ubiquitination regulates the internalization efficiency and might help in promoting internalization of the desensitized MOP.
Insights
Ubiquitination is not required for Gi/o signaling or opioid receptor phosphorylation. However, this modification is crucial for efficient internalization of the desensitized μ-opioid receptor (MOP).
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Opioids are potent analgesics that induce tolerance.
- Phosphorylation of the μ-opioid receptor (MOP) is key to receptor desensitization and cellular sensitivity.
- Ubiquitination, a posttranslational modification, is increasingly recognized for its role in receptor signaling and regulation.
Purpose of the Study:
- To investigate the role of MOP ubiquitination in Gi/o-mediated signaling, desensitization, and internalization.
- To determine if ubiquitination is essential for opioid-induced cellular responses.
Main Methods:
- Generation of a ubiquitination-deficient MOP mutant.
- Assays to measure Gi/o pathway activation.
- Analysis of receptor phosphorylation.
- Assessment of receptor desensitization and internalization.
Main Results:
- Gi/o-mediated signaling and MOP phosphorylation proceed independently of ubiquitination.
- Ubiquitination is not required for receptor desensitization.
- Ubiquitination significantly enhances the efficiency of MOP internalization, particularly for desensitized receptors.
Conclusions:
- Ubiquitination is a critical regulator of μ-opioid receptor internalization, not its initial signaling or desensitization.
- Targeting MOP ubiquitination may offer novel strategies for modulating opioid efficacy and managing tolerance.
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