Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by regulators of G protein signaling proteins 4 or 8 and 7 in permeabilised C6 cells is Gα subtype dependent

Jeffery N TALBOT; David L ROMAN; Mary J CLARK; Rebecca A ROOF; John J. G TESMER; Richard R NEUBIG; John R TRAYNOR

doi:10.1111/j.1471-4159.2009.06519.x

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Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by regulators of G protein signaling proteins 4 or 8 and 7 in permeabilised C6 cells is Gα subtype dependent

Journal article

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Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by regulators of G protein signaling proteins 4 or 8 and 7 in permeabilised C6 cells is Gα subtype dependent

Jeffery N TALBOT, David L ROMAN, Mary J CLARK, Rebecca A ROOF, John J. G TESMER, Richard R NEUBIG and John R TRAYNOR

Journal of neurochemistry, Vol.112(4), pp.1026-1034

2010

DOI: 10.1111/j.1471-4159.2009.06519.x

PMID: 20002516

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Abstract

Regulators of G protein signaling (RGS) proteins act as GTPase-accelerating protein to negatively modulate G protein signaling and are defined by a conserved RGS domain with considerable amino acid diversity. To determine the effects of specific, purified RGS proteins on mu-opioid signaling, C6 cells stably expressing a mu-opioid receptor were rendered permeable to proteins by treatment with digitonin. Mu-opioid inhibition of forskolin-stimulated adenylyl cyclase by [d-Ala2,N-Me-Phe4,Gly-ol]-enkephalin (DAMGO), a mu-specific opioid peptide, remained fully intact in permeabilized cells. Purified RGS domain of RGS4 added to permeabilized cells resulted in a twofold loss in DAMGO potency but had no effect in cells expressing RGS-insensitive G proteins. The inhibitory effect of DAMGO was reduced to the same extent by purified RGS4 and RGS8. In contrast, the RGS domain of RGS7 had no effect and inhibited the action of RGS8 as a result of weak physical association with Gαi2 and minimal GTPase-accelerating protein activity in C6 cell membranes. These data suggest that differences in conserved RGS domains of specific RGS proteins contribute to differential regulation of opioid signaling to adenylyl cyclase and that a permeabilized cell model is useful for studying the effects of specific RGS proteins on aspects of G protein-coupled receptor signaling.

Cell Physiology

Signal Transduction

Fundamental and applied biological sciences. Psychology

Cell receptors

Neuropeptide receptors

Biological and medical sciences

Molecular and cellular biology

Cell structures and functions

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Title: Subtitle: Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by regulators of G protein signaling proteins 4 or 8 and 7 in permeabilised C6 cells is Gα subtype dependent
Creators: Jeffery N TALBOT - Department of Pharmacology, University of Michigan, Ann Arbor, MI, United States
David L ROMAN - Department of Pharmacology, University of Michigan, Ann Arbor, MI, United States
Mary J CLARK - Department of Pharmacology, University of Michigan, Ann Arbor, MI, United States
Rebecca A ROOF - Department of Pharmacology, University of Michigan, Ann Arbor, MI, United States
John J. G TESMER - Department of Pharmacology, University of Michigan, Ann Arbor, MI, United States
Richard R NEUBIG - Department of Pharmacology, University of Michigan, Ann Arbor, MI, United States
John R TRAYNOR - Department of Pharmacology, University of Michigan, Ann Arbor, MI, United States
Resource Type: Journal article
Publication Details: Journal of neurochemistry, Vol.112(4), pp.1026-1034
Publisher: Wiley-Blackwell; Oxford
DOI: 10.1111/j.1471-4159.2009.06519.x
PMID: 20002516
ISSN: 0022-3042
eISSN: 1471-4159
Language: English
Date published: 2010
Academic Unit: Pharmacy; Iowa Neuroscience Institute; Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
Record Identifier: 9984065484702771

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