Analysis of selective binding epitopes for the κ-opioid receptor antagonist nor-binaltorphimine

S. A. Hjorth, K. Thirstrup, David Grandy, T. W. Schwartz

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Abstract

The structural determinants for the selective binding of the nonpeptide opioid receptor antagonist nor-binaltorphimine (nor-BNI) to the ≃-opioid receptor were characterized using a systematic series of chimeras between the ≃ receptor and the homologous μ-opioid receptor. All 10 chimeric constructs bound the nonselective antagonists (-)-naloxone and diprenorphine with similar affinities, as did the two wild-type receptors. Introduction of amino-terminal segments of increasing length, extending to and including transmembrane segment VI, from the μ receptor into the ≃ receptor did not impair the high affinity binding of nor-BNI, and neither did introduction of the intracellular carboxyl-terminal extension of the μ receptor. In contrast, nor-BNI binding was impaired ≥600-fold in constructs in which extracellular loop 3 and transmembrane segment VII originated from the μ receptor. The exchange of a single residue within this region, Glu297, for lysine, the corresponding residue from the μ receptor, reduced the binding affinity of nor-BNI 142-fold, without affecting the binding of the nonselective compounds (-)-naloxone and diprenorphine. It is concluded that the selective binding of nor-BNI to the ≃-opioid receptor is determined by nonconserved residues located in extracellular loop 3 and transmembrane segment VII and that Glu297, located just outside transmembrane segment VI, plays a major role in the ≃-selective binding characteristics of nor- BNI.

Original languageEnglish (US)
Pages (from-to)1089-1094
Number of pages6
JournalMolecular Pharmacology
Volume47
Issue number6
StatePublished - 1995
Externally publishedYes

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Narcotic Antagonists
Epitopes
Opioid Receptors
Diprenorphine
Naloxone
Lysine
norbinaltorphimine

ASJC Scopus subject areas

  • Pharmacology

Cite this

Hjorth, S. A., Thirstrup, K., Grandy, D., & Schwartz, T. W. (1995). Analysis of selective binding epitopes for the κ-opioid receptor antagonist nor-binaltorphimine. Molecular Pharmacology, 47(6), 1089-1094.

Analysis of selective binding epitopes for the κ-opioid receptor antagonist nor-binaltorphimine. / Hjorth, S. A.; Thirstrup, K.; Grandy, David; Schwartz, T. W.

In: Molecular Pharmacology, Vol. 47, No. 6, 1995, p. 1089-1094.

Research output: Contribution to journalArticle

Hjorth, SA, Thirstrup, K, Grandy, D & Schwartz, TW 1995, 'Analysis of selective binding epitopes for the κ-opioid receptor antagonist nor-binaltorphimine', Molecular Pharmacology, vol. 47, no. 6, pp. 1089-1094.
Hjorth, S. A. ; Thirstrup, K. ; Grandy, David ; Schwartz, T. W. / Analysis of selective binding epitopes for the κ-opioid receptor antagonist nor-binaltorphimine. In: Molecular Pharmacology. 1995 ; Vol. 47, No. 6. pp. 1089-1094.
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