Properties of a κ-opioid receptor expressed in CHO cells: interaction with multiple G-proteins is not specific for any individual Gα subunit and is similar to that of other opioid receptors

P. L. Prather, T. M. McGinn, P. A. Claude, L. Y. Liu-Chen, Horace H Loh, Ping-Yee Law

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Abstract

The purpose of the present study was to examine the coupling pattern of a recently cloned κ-opioid receptor stably transfected in CHO cells to individual Gα subunits with subsequent comparison to that observed previously for δ- and μ-opioid receptors. Data presented in the current study indicate the successful stable expression of a κ-opioid receptor in CHO cells. This is supported by experiments in which ligands with selectivity for κ-, but not δ- or μ-opioid receptors demonstrated high affinity for the expressed receptor and were able to potently and efficaciously produce inhibition of adenylyl cyclase activity. In addition, only κ-opioid agonists were able to induce dose-dependent increases in the incorporation of [32P]azidoanilido-GTP into four Gα subunits, three of which were identified as Gi, Gi and Go,. Further, the amount of K-opioid agonists required to induce 50% maximal labeling of any individual Gα subunit was similar. Although κ-opioid agonists produced equivalent maximal labeling of Gi, Gi, and Go, significantly less agonist-induced labeling was observed for an unknown G-protein designated as G?α. Although these results are slightly different than those observed previously for both δ- and μ-opioid receptors, it appears that all opioid receptors stably transfected in CHO cells interact with multiple G-proteins and that this coupling is not selective for any invidivual Gα subunit.

Original languageEnglish (US)
Pages (from-to)336-346
Number of pages11
JournalMolecular Brain Research
Volume29
Issue number2
DOIs
StatePublished - Apr 1995

Bibliographical note

Funding Information:
This research was supported in part by NIDA Grants no. DA07339, no. DA05695, and NIDA training Grant no. DA07234-07.

Keywords

  • Adenylyl cyclase
  • CHO cell
  • Dynorphin-A(1-17)
  • G-protein
  • Transfection
  • U50-488H
  • κ-Opioid receptor

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