Guanine nucleotide-mediated inhibition of opioid agonist binding: Modulatory effects of ions and of receptor occupancy

doi:10.1016/0006-2952(89)90491-7

Biochemical Pharmacology

Volume 38, Issue 12, 15 June 1989, Pages 1931-1939

https://doi.org/10.1016/0006-2952(89)90491-7 Get rights and content

Abstract

We have analysed the potency of GTP, GDP and their analogues in reducing [³H]DADLE binding to opioid receptors in NG 108-15 cell membranes. Under conditions where non-specific hydrolysis and transphosphorylation is inhibited, the following rank order of potency was found: GDP ≥ GTPγS > GTP / GDPβS ≥ GDPNH₂ > GppNHp / GMP. Remarkably, the slopes for the inhibition curves of GTP, GDP and their thiosubstituted analogues, but not of GDPNH₂ and GppNHp, were extremely shallow, indicating either negative cooperativity or the existence of two states for the guanine nucleotide binding proteins, that both can mediate the effect of nucleotides on agonist receptor binding. The potencies of the different guanine nucleotide analogues, except that of GppNHp, were increased by the presence of sodium or chloride ions in the assay medium. Magnesium also affected GTP-mediated inhibition of opioid agonist binding since it decreased the ic₅₀ of the nucleotide and steepened the slope of the inhibition curve. The ic₅₀s of nucleotides and the slopes of their inhibition curves were also dependent on the extent of receptor occupancy by the agonist. From these data we conclude that (1) either diphospho- or triphosphonucleotides can regulate agonist binding. (2) Magnesium, sodium and chloride, by acting at different components of the receptorJG protein complex produce similar effects on nucleotide mediated regulation of agonist binding. (3) A mutual influence exists between receptor occupancy by agonists and G protein-mediated guanine nucleotide effect on the receptor.

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^∗: Present address: Department of Biopharmacy, Institute of Pharmacy, ETHZ, Clausiusstrasse 25, CH-8092 Zürich, Switzerland.

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Guanine nucleotide-mediated inhibition of opioid agonist binding: Modulatory effects of ions and of receptor occupancy

Abstract

J Biol Chem

J Biol Chem

J Biol Chem

J Biol Chem

J Biol Chem

Methods Enzymol

J Biol Chem

Anal Biochem

J Biol Chem

J Biol Chem

J Biol Chem

J Biol Chem

J Biol Chem

J Bio Chem

J Biol Chem

G proteins: a family of signal transducers

Ann Rev Cell Biol

The role of hormone receptors and GTP regulatory proteins in membrane transduction

Nature

G proteins: transducers of receptor-generated signals

Ann Rev Biochem