Receptor binding of [3H]naloxone benzoylhydrazone

A reversible κ and slowly dissociable μ opiate

M. Price, M. A. Gistrak, Yossef Itzhak, E. F. Hahn, G. W. Pasternak

Research output: Contribution to journalArticle

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Abstract

In standard 3H-opioid binding assays, the benzoylhydrazone derivative of naloxone (6-desoxy-6-benzoylhydrazido-N-allyl-14-hydroxydihydronormorphinone; Nh alBzoH) inhibited μ, κ, and δ binding at nanomolar concentrations. At concentrations as low as 1 nM, it also produced a wash-resistant inhibition of opioid binding. [3H]NalBzoH binding typically gave a ratio of total to nonspecific binding of 8:1. Binding reached state levels by 1 hr and was linear with tissue concentration. [3H]NalBzoH labeled two classes of sites. The binding to one was easily reversible whereas the other was not and was termed pseudoir-reversible. At 25°, almost 90% of [3H]naloxone binding and approximately 60-75% of [3H]NalBzoH binding dissociated over 90 min. However, the remainder of [3H]NalBzoH binding, corresponding to pseudoirreversible binding, remained constant over the next 5 hr at 25° and additional studies suggested a dissociation half-life of approximately 24 hr. Competition studies indicated that the reversible binding corresponded to neither μ nor δ binding and may represent a novel subtype of κ receptor. Pseudoirreversible binding was predominantly to a combination of both μ1 and μ2 receptors. Despite its extremely slow rate of dissociation, pseudoirreversible binding was not covalent inasmuch as lowering the pH to 5 or adding the GTP analog 5'-guanylylimidodiphosphate [Gpp(NH)p] completely dissociated prebound [3H]NalBzoH. The ability of Gpp(NH)p to dissociate pseudoirreversible [3H]NalBsoH binding raised the possibility that the slow rate of dissociation was related to interactions with a guanine nucleotide-binding protein.

Original languageEnglish
Pages (from-to)67-74
Number of pages8
JournalMolecular Pharmacology
Volume35
Issue number1
StatePublished - Jan 1 1989
Externally publishedYes

Fingerprint

Opiate Alkaloids
Guanylyl Imidodiphosphate
Opioid Analgesics
Guanine Nucleotides
Naloxone
Guanosine Triphosphate
naloxone benzoylhydrazone
Half-Life
Carrier Proteins

ASJC Scopus subject areas

  • Pharmacology

Cite this

Price, M., Gistrak, M. A., Itzhak, Y., Hahn, E. F., & Pasternak, G. W. (1989). Receptor binding of [3H]naloxone benzoylhydrazone: A reversible κ and slowly dissociable μ opiate. Molecular Pharmacology, 35(1), 67-74.

Receptor binding of [3H]naloxone benzoylhydrazone : A reversible κ and slowly dissociable μ opiate. / Price, M.; Gistrak, M. A.; Itzhak, Yossef; Hahn, E. F.; Pasternak, G. W.

In: Molecular Pharmacology, Vol. 35, No. 1, 01.01.1989, p. 67-74.

Research output: Contribution to journalArticle

Price, M, Gistrak, MA, Itzhak, Y, Hahn, EF & Pasternak, GW 1989, 'Receptor binding of [3H]naloxone benzoylhydrazone: A reversible κ and slowly dissociable μ opiate', Molecular Pharmacology, vol. 35, no. 1, pp. 67-74.
Price M, Gistrak MA, Itzhak Y, Hahn EF, Pasternak GW. Receptor binding of [3H]naloxone benzoylhydrazone: A reversible κ and slowly dissociable μ opiate. Molecular Pharmacology. 1989 Jan 1;35(1):67-74.
Price, M. ; Gistrak, M. A. ; Itzhak, Yossef ; Hahn, E. F. ; Pasternak, G. W. / Receptor binding of [3H]naloxone benzoylhydrazone : A reversible κ and slowly dissociable μ opiate. In: Molecular Pharmacology. 1989 ; Vol. 35, No. 1. pp. 67-74.
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