Plasma membrane localization of the μ-opioid receptor controls spatiotemporal signaling

Halls, M.L.; Yeatman, H.R.; Nowell, C.J.; Thompson, G.L.; Gondin, A.B.; Civciristov, S.; Bunnett, N.W.; Lambert, N.A.; Poole, D.P.; Canals, M.

Abstract

Differential regulation of the μ-opioid receptor (MOR), a G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptor, contributes to the clinically limiting effects of opioid analgesics, such as morphine. We used biophysical approaches to quantify spatiotemporal MOR signaling in response to different ligands. In human embryonic kidney (HEK) 293 cells over-expressing MOR, morphine caused a Gβγ-dependent increase in plasma membrane-localized protein kinase C (PKC) activity, which resulted in a restricted distribution of MOR within the plasma membrane and induced sustained cytosolic extracellular signal-regulated kinase (ERK) signaling. In contrast, the synthetic opioid peptide DAMGO ([D-Ala2,N-Me-Phe4, Gly5-ol]-enkephalin) enabled receptor redistribution within the plasma membrane, resulting in transient increases in cytosolic and nuclear ERK activity, and, subsequently, receptor internalization. When Gβγ subunits or PKCα activity was inhibited or when the carboxyl-terminal phosphorylation sites of MOR were mutated, morphine-activated MOR was released from its restricted plasma membrane localization and stimulated a transient increase in cytosolic and nuclear ERK activity in the absence of receptor internalization. Thus, these data suggest that the ligand-induced redistribution of MOR within the plasma membrane, and not its internalization, controls its spatiotemporal signaling.

Citation

Halls, M., Yeatman, H., Nowell, C., Thompson, G., Gondin, A., Civciristov, S., …Canals, M. (2016). Plasma membrane localization of the μ-opioid receptor controls spatiotemporal signaling. Science Signaling, 9(414), ra16-ra16. https://doi.org/10.1126/scisignal.aac9177

• Journal Article Type: Article
• Acceptance Date: Jan 25, 2016
• Online Publication Date: Feb 9, 2016
• Publication Date: Feb 9, 2016
• Deposit Date: Mar 6, 2019
• Publicly Available Date: Mar 6, 2019
• Journal: Science Signaling
• Print ISSN: 1945-0877
• Electronic ISSN: 1937-9145
• Publisher: American Association for the Advancement of Science
• Peer Reviewed: Peer Reviewed
• Volume: 9
• Issue: 414
• Pages: ra16-ra16
• DOI: https://doi.org/10.1126/scisignal.aac9177
• Keywords: enkephalin[2 dextro alanine 4 methylphenylalanine 5 glycine]; G beta gamma complex; guanine nucleotide binding protein; mitogen activated protein kinase; morphine; mu opiate receptor; protein kinase C; unclassified drug; enkephalin[2 dextro alanine 4 meth
• Public URL: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84958531919&doi=10.1126%2fscisignal.aac9177&partnerID=40&md5=2c0e1199b3a7fd3d8055e6cde401aea9
• Publisher URL: http://stke.sciencemag.org/content/9/414/ra16.long
• Additional Information: This is the author’s version of the work. It is posted here by permission of the AAAS for personal use, not for redistribution. The definitive version was published in Science Signaling on 09 Feb 2016, vol. 9, issue 414, DOI: 10.1126/scisignal.aac9177 .