Molecular cloning and functional expression of a μ-opioid receptor from rat brain

Yan Chen, Anton Mestek, Jian Liu, Joyce Hurley, Lei Yu

Research output: Contribution to journalArticle

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Abstract

Opioid drugs act on specific receptors to modulate a wide range of physiological functions. There are at least three types of opioid receptors, μ, δ, and κ. Using a cDNA probe for a mouse γ-opioid receptor in low stringency hybridization, a clone has been isolated from a rat brain cDNA library. This clone contains an open reading frame of 1194 base pairs, with a deduced polypeptide of 398 amino acid residues. The predicted protein exhibits the structural features of guanine nucleotide-binding protein-coupled receptors and displays a high degree of sequence homology with the mouse γ-opioid receptor. When transfected into COS-7 cells, the cDNA conferred a binding site with subnanomolar affinity for [3H]diprenorphine, a high affinity ligand for all three types of opioid receptors. This site also displayed nanomolar affinity for [D-Ala2,N-Me-Phe4,Gly-ol5]-enkephalin (DAGO), a μ-selective agonist, whereas its affinities for the γ-selective agonist [D-Pen2,5]-enkephalin and the κ-selective agonist U-50488 were in the micromolar range. Several μ-selective antagonists, including naloxonazine, β-funaltrexamine, and cyprodime, were capable of displacing [3H]diprenorphine binding with nanomolar potency. The pharmacological profile of this binding site thus suggests that it is a μ-type opioid receptor, which we designated MOR-1. In COS-7 cells expressing MOR-1 and stimulated with forskolin, treatment with DAGO decreased the steady state levels of cAMP; this inhibitory effect of DAGO was blocked by naloxonazine. These results suggest that this μ-opioid receptor is functionally coupled to the inhibition of adenylyl cyclase.

Original languageEnglish
Pages (from-to)8-12
Number of pages5
JournalMolecular Pharmacology
Volume44
Issue number1
StatePublished - Jul 1993

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Molecular Cloning
Opioid Receptors
Ala(2)-MePhe(4)-Gly(5)-enkephalin
Brain
Diprenorphine
Enkephalins
COS Cells
Complementary DNA
Clone Cells
Binding Sites
(trans)-Isomer 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide
Guanine Nucleotides
Colforsin
Sequence Homology
Gene Library
Adenylyl Cyclases
Base Pairing
Opioid Analgesics
Open Reading Frames
Carrier Proteins

ASJC Scopus subject areas

  • Pharmacology

Cite this

Molecular cloning and functional expression of a μ-opioid receptor from rat brain. / Chen, Yan; Mestek, Anton; Liu, Jian; Hurley, Joyce; Yu, Lei.

In: Molecular Pharmacology, Vol. 44, No. 1, 07.1993, p. 8-12.

Research output: Contribution to journalArticle

Chen, Yan ; Mestek, Anton ; Liu, Jian ; Hurley, Joyce ; Yu, Lei. / Molecular cloning and functional expression of a μ-opioid receptor from rat brain. In: Molecular Pharmacology. 1993 ; Vol. 44, No. 1. pp. 8-12.
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