Allosterism within d opioid-k opioid receptor heteromers in peripheral sensory neurons: Regulation of k opioid agonist efficacy

Blaine A. Jacobs, Miryam M. Pando, Elaine Jennings, Teresa A. Chavera, William P Clarke, Kelly A Berg

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Abstract

There is abundant evidence for formation of G protein-coupled receptor heteromers in heterologous expression systems, but little is known of the function of heteromers in native systems. Heteromers of d and k opioid receptors (DOR-KOR heteromers) have been identified in native systems. We previously reported that activation of DOR-KOR heteromers expressed by rat painsensing neurons (nociceptors) produces robust, peripherally mediated antinociception. Moreover, DOR agonist potency and efficacy is regulated by KOR antagonists via allosteric interactions within the DOR-KOR heteromer in a ligand-dependent manner. Here we assessed the reciprocal regulation of KOR agonist function by DOR antagonists in adult rat nociceptors in culture and in a behavioral assay of nociception. Naltrindole enhanced the potency of the KOR agonist 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-phenyl-2-pyrrolidin-1-ylethyl]acetamide (ICI-199441) 10-to 20-fold, but did not alter responses to 2-(3,4-dichlorophenyl)-N-methyl-N-[(1R,2R)-2-pyrrolidin-1-ylcyclohexyl]acetamide (U50488). By contrast, the potency of U50488 was enhanced 20-fold by 7-benzylidenenaltrexone. The efficacy of 6′-guanidinonaltrindole (6′-GNTI) to inhibit nociceptors was blocked by small interfering RNA knockdown of DOR or KOR. Replacing 6′-GNTI occupancy of DOR with either naltrindole or 7-benzylidenenaltrexone abolished 6′-GNTI efficacy. Further, peptides derived from DOR transmembrane segment 1 fused to the cell membrane-penetrating HIV transactivator of transcription peptide also blocked 6′-GNTI-mediated responses ex vivo and in vivo, suggesting that 6′-GNTI efficacy in nociceptors is due to its positive allosteric regulation of KOR via occupancy of DOR in a DOR-KOR heteromer. Together, these results provide evidence for the existence of functional DOR-KOR heteromers in rat peripheral sensory neurons and that reciprocal, ligand-dependent allosteric interactions occur between the DOR and KOR protomers.

Original languageEnglish (US)
Pages (from-to)376-386
Number of pages11
JournalMolecular Pharmacology
Volume93
Issue number4
DOIs
StatePublished - Apr 1 2018

Fingerprint

Opioid Receptors
Sensory Receptor Cells
Nociceptors
naltrindole
Opioid Analgesics
Allosteric Regulation
Ligands
Peptides
Nociception
Trans-Activators
Protein Subunits
G-Protein-Coupled Receptors
Small Interfering RNA
Cell Membrane
6'-guanidinonaltrindole
HIV
Neurons
7-benzylidenenaltrexone
acetamide

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

Allosterism within d opioid-k opioid receptor heteromers in peripheral sensory neurons : Regulation of k opioid agonist efficacy. / Jacobs, Blaine A.; Pando, Miryam M.; Jennings, Elaine; Chavera, Teresa A.; Clarke, William P; Berg, Kelly A.

In: Molecular Pharmacology, Vol. 93, No. 4, 01.04.2018, p. 376-386.

Research output: Contribution to journalArticle

Jacobs, Blaine A. ; Pando, Miryam M. ; Jennings, Elaine ; Chavera, Teresa A. ; Clarke, William P ; Berg, Kelly A. / Allosterism within d opioid-k opioid receptor heteromers in peripheral sensory neurons : Regulation of k opioid agonist efficacy. In: Molecular Pharmacology. 2018 ; Vol. 93, No. 4. pp. 376-386.
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