Multiple affinity binding states of the σ receptor

Effect of GTP-binding protein-modifying agents

Yossef Itzhak

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

The σ receptor, which is labeled with (+)-[3H]3-(3-hydroxyphenyl)-N-1-(propyl)piperidine [(+)-[3H]3-PPP], is a site that binds several psychotomimetic opiate benzomorphans and certain antipsychotics, such as haloperidol. In order to elucidate the mechanisms involved in σ receptor ligand binding, equilibrium binding analysis and kinetics of association and dissociation of the relatively selective σ receptor ligand (+)-[3H]3-PPP were determined in rat brain membranes in the absence and presence of 5'-guanylylimidodiphosphate [Gpp(NH)p]. In the absence of Gpp(NH)p, (+)-3-PPP, cyclazocine, pentazocine, and (+)-SKF 10047 bind to high and low affinity sites (K(H) = 1.3-7.5 nM; K(L) = 84-500 nM), as determined by computer assisted analysis of the inhibition of (+)-[3H]3-PPP binding by the σ ligands. The antipsychotics haloperidol and chlorpromazine inhibit (+)-[3H]3-PPP binding in a manner indicating interaction with a single state of the receptor. Gpp(NH)p (0.1 mM) abolished the high affinity binding component of the σ agonist-like compounds tested but had no effect on the affinities of the antipsychotics for the receptor. Gpp(NH)p decreased the association rate of (+)-[3H]3-PPP binding 5-fold and also converted the biexponential dissociation kinetics of the ligand, observed in the absence of Gpp(NH)p, to a rapid monophasic dissociation process. Pretreatment of membranes with N-ethylmaleimide and pertussis toxin inhibited (+)-[3H]3-PPP binding and abolished the effect of Gpp(NH)p on the σ ligand binding. These findings indicate that the σ receptor is capable of existing in two discrete states, having high and low affinity for σ agonist-like drugs. The regulation of the high affinity binding state by GTP-binding protein-modifying agents suggests its coupling to GTP-binding protein(s).

Original languageEnglish
Pages (from-to)512-517
Number of pages6
JournalMolecular Pharmacology
Volume36
Issue number4
StatePublished - Jan 1 1989

Fingerprint

Guanylyl Imidodiphosphate
GTP-Binding Proteins
Ligands
Antipsychotic Agents
Haloperidol
Opiate Alkaloids
Benzomorphans
Cyclazocine
Pentazocine
Ethylmaleimide
Membranes
Pertussis Toxin
Chlorpromazine
preclamol
Brain
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Pharmacology

Cite this

Multiple affinity binding states of the σ receptor : Effect of GTP-binding protein-modifying agents. / Itzhak, Yossef.

In: Molecular Pharmacology, Vol. 36, No. 4, 01.01.1989, p. 512-517.

Research output: Contribution to journalArticle

@article{12f37a2125b14884a3680d8a300214f3,
title = "Multiple affinity binding states of the σ receptor: Effect of GTP-binding protein-modifying agents",
abstract = "The σ receptor, which is labeled with (+)-[3H]3-(3-hydroxyphenyl)-N-1-(propyl)piperidine [(+)-[3H]3-PPP], is a site that binds several psychotomimetic opiate benzomorphans and certain antipsychotics, such as haloperidol. In order to elucidate the mechanisms involved in σ receptor ligand binding, equilibrium binding analysis and kinetics of association and dissociation of the relatively selective σ receptor ligand (+)-[3H]3-PPP were determined in rat brain membranes in the absence and presence of 5'-guanylylimidodiphosphate [Gpp(NH)p]. In the absence of Gpp(NH)p, (+)-3-PPP, cyclazocine, pentazocine, and (+)-SKF 10047 bind to high and low affinity sites (K(H) = 1.3-7.5 nM; K(L) = 84-500 nM), as determined by computer assisted analysis of the inhibition of (+)-[3H]3-PPP binding by the σ ligands. The antipsychotics haloperidol and chlorpromazine inhibit (+)-[3H]3-PPP binding in a manner indicating interaction with a single state of the receptor. Gpp(NH)p (0.1 mM) abolished the high affinity binding component of the σ agonist-like compounds tested but had no effect on the affinities of the antipsychotics for the receptor. Gpp(NH)p decreased the association rate of (+)-[3H]3-PPP binding 5-fold and also converted the biexponential dissociation kinetics of the ligand, observed in the absence of Gpp(NH)p, to a rapid monophasic dissociation process. Pretreatment of membranes with N-ethylmaleimide and pertussis toxin inhibited (+)-[3H]3-PPP binding and abolished the effect of Gpp(NH)p on the σ ligand binding. These findings indicate that the σ receptor is capable of existing in two discrete states, having high and low affinity for σ agonist-like drugs. The regulation of the high affinity binding state by GTP-binding protein-modifying agents suggests its coupling to GTP-binding protein(s).",
author = "Yossef Itzhak",
year = "1989",
month = "1",
day = "1",
language = "English",
volume = "36",
pages = "512--517",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "4",

}

TY - JOUR

T1 - Multiple affinity binding states of the σ receptor

T2 - Effect of GTP-binding protein-modifying agents

AU - Itzhak, Yossef

PY - 1989/1/1

Y1 - 1989/1/1

N2 - The σ receptor, which is labeled with (+)-[3H]3-(3-hydroxyphenyl)-N-1-(propyl)piperidine [(+)-[3H]3-PPP], is a site that binds several psychotomimetic opiate benzomorphans and certain antipsychotics, such as haloperidol. In order to elucidate the mechanisms involved in σ receptor ligand binding, equilibrium binding analysis and kinetics of association and dissociation of the relatively selective σ receptor ligand (+)-[3H]3-PPP were determined in rat brain membranes in the absence and presence of 5'-guanylylimidodiphosphate [Gpp(NH)p]. In the absence of Gpp(NH)p, (+)-3-PPP, cyclazocine, pentazocine, and (+)-SKF 10047 bind to high and low affinity sites (K(H) = 1.3-7.5 nM; K(L) = 84-500 nM), as determined by computer assisted analysis of the inhibition of (+)-[3H]3-PPP binding by the σ ligands. The antipsychotics haloperidol and chlorpromazine inhibit (+)-[3H]3-PPP binding in a manner indicating interaction with a single state of the receptor. Gpp(NH)p (0.1 mM) abolished the high affinity binding component of the σ agonist-like compounds tested but had no effect on the affinities of the antipsychotics for the receptor. Gpp(NH)p decreased the association rate of (+)-[3H]3-PPP binding 5-fold and also converted the biexponential dissociation kinetics of the ligand, observed in the absence of Gpp(NH)p, to a rapid monophasic dissociation process. Pretreatment of membranes with N-ethylmaleimide and pertussis toxin inhibited (+)-[3H]3-PPP binding and abolished the effect of Gpp(NH)p on the σ ligand binding. These findings indicate that the σ receptor is capable of existing in two discrete states, having high and low affinity for σ agonist-like drugs. The regulation of the high affinity binding state by GTP-binding protein-modifying agents suggests its coupling to GTP-binding protein(s).

AB - The σ receptor, which is labeled with (+)-[3H]3-(3-hydroxyphenyl)-N-1-(propyl)piperidine [(+)-[3H]3-PPP], is a site that binds several psychotomimetic opiate benzomorphans and certain antipsychotics, such as haloperidol. In order to elucidate the mechanisms involved in σ receptor ligand binding, equilibrium binding analysis and kinetics of association and dissociation of the relatively selective σ receptor ligand (+)-[3H]3-PPP were determined in rat brain membranes in the absence and presence of 5'-guanylylimidodiphosphate [Gpp(NH)p]. In the absence of Gpp(NH)p, (+)-3-PPP, cyclazocine, pentazocine, and (+)-SKF 10047 bind to high and low affinity sites (K(H) = 1.3-7.5 nM; K(L) = 84-500 nM), as determined by computer assisted analysis of the inhibition of (+)-[3H]3-PPP binding by the σ ligands. The antipsychotics haloperidol and chlorpromazine inhibit (+)-[3H]3-PPP binding in a manner indicating interaction with a single state of the receptor. Gpp(NH)p (0.1 mM) abolished the high affinity binding component of the σ agonist-like compounds tested but had no effect on the affinities of the antipsychotics for the receptor. Gpp(NH)p decreased the association rate of (+)-[3H]3-PPP binding 5-fold and also converted the biexponential dissociation kinetics of the ligand, observed in the absence of Gpp(NH)p, to a rapid monophasic dissociation process. Pretreatment of membranes with N-ethylmaleimide and pertussis toxin inhibited (+)-[3H]3-PPP binding and abolished the effect of Gpp(NH)p on the σ ligand binding. These findings indicate that the σ receptor is capable of existing in two discrete states, having high and low affinity for σ agonist-like drugs. The regulation of the high affinity binding state by GTP-binding protein-modifying agents suggests its coupling to GTP-binding protein(s).

UR - http://www.scopus.com/inward/record.url?scp=0024433774&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024433774&partnerID=8YFLogxK

M3 - Article

VL - 36

SP - 512

EP - 517

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 4

ER -