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Title: Solubilization of a guanyl nucleotide-sensitive alpha/sub 1/ adrenergic receptor from liver membranes

Abstract

Rat liver membranes incubated with norepinephrine before solubilization with digitonin yielded a soluble hormone-receptor complex from which the release of tightly bound norepinephrine was facilitated by guanyl nucleotides. Binding of the alpha/sub 1/-adrenergic receptor antagonist, (/sup 3/H)-prazosin, to the soluble preparation was utilized as a gauge of guanyl nucleotide-induced release of receptor-bound agonist. The following potency series was obtained with regard to the ability of guanyl nucleotides to facilitate (/sup 3/H)-prazosin binding to the solubilized preparation: guanosine 5'-0-(3-thiotriphosphate)(K/sub 1/2/ = 2.5 nM), guanylyl-imidodiphosphate (K/sub 1/2/ = 10 nM), guanosine triphosphate (K/sub 1/2/ = 34 nM) and adenylyl-imidodiphosphate (K/sub 1/2/ > 1 mM). In the presence of guanylyl-imidodiphosphate (0.4 mM), the receptor population displayed monotonic binding parameters with a K/sub d/ for (/sup 3/H)-prazosin of 1.16 nM by Scatchard analysis. Competition curves against (/sup 3/H)-prazosin with the antagonists phentolamine and yohimbine revealed respective K/sub i/'s of .089 ..mu..M and 1.8 ..mu..M; curves with the agonists norepinephrine and isoproterenol yielded respective K/sub i/'s of 6.2..mu..M and 360 ..mu..M. Competition curves performed in the absence of guanyl nucleotide were complex demonstrating an apparent increase in affinity for agonists and an apparent decrease in affinity for antagonists. These curve shifts are consistent with themore » conversion of receptor to and from the guanyl nucleotide-sensitive state as a function of competing ligand concentration.« less

Authors:
;
Publication Date:
Research Org.:
National Institute of Health, Bethesda, MD
OSTI Identifier:
5904399
Report Number(s):
CONF-870644-
Journal ID: CODEN: FEPRA; TRN: 87-037163
Resource Type:
Conference
Journal Name:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
Additional Journal Information:
Journal Volume: 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; RECEPTORS; BIOCHEMICAL REACTION KINETICS; SOLUBILITY; SYMPATHOMIMETICS; AFFINITY; CELL MEMBRANES; LIGANDS; LIVER; NUCLEOTIDES; RATS; TRACER TECHNIQUES; TRITIUM COMPOUNDS; ANIMALS; AUTONOMIC NERVOUS SYSTEM AGENTS; BODY; CELL CONSTITUENTS; DIGESTIVE SYSTEM; DRUGS; GLANDS; ISOTOPE APPLICATIONS; KINETICS; LABELLED COMPOUNDS; MAMMALS; MEMBRANE PROTEINS; MEMBRANES; ORGANIC COMPOUNDS; ORGANS; PROTEINS; REACTION KINETICS; RODENTS; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Harris, S I, and Moss, J. Solubilization of a guanyl nucleotide-sensitive alpha/sub 1/ adrenergic receptor from liver membranes. United States: N. p., 1987. Web.
Harris, S I, & Moss, J. Solubilization of a guanyl nucleotide-sensitive alpha/sub 1/ adrenergic receptor from liver membranes. United States.
Harris, S I, and Moss, J. 1987. "Solubilization of a guanyl nucleotide-sensitive alpha/sub 1/ adrenergic receptor from liver membranes". United States.
@article{osti_5904399,
title = {Solubilization of a guanyl nucleotide-sensitive alpha/sub 1/ adrenergic receptor from liver membranes},
author = {Harris, S I and Moss, J},
abstractNote = {Rat liver membranes incubated with norepinephrine before solubilization with digitonin yielded a soluble hormone-receptor complex from which the release of tightly bound norepinephrine was facilitated by guanyl nucleotides. Binding of the alpha/sub 1/-adrenergic receptor antagonist, (/sup 3/H)-prazosin, to the soluble preparation was utilized as a gauge of guanyl nucleotide-induced release of receptor-bound agonist. The following potency series was obtained with regard to the ability of guanyl nucleotides to facilitate (/sup 3/H)-prazosin binding to the solubilized preparation: guanosine 5'-0-(3-thiotriphosphate)(K/sub 1/2/ = 2.5 nM), guanylyl-imidodiphosphate (K/sub 1/2/ = 10 nM), guanosine triphosphate (K/sub 1/2/ = 34 nM) and adenylyl-imidodiphosphate (K/sub 1/2/ > 1 mM). In the presence of guanylyl-imidodiphosphate (0.4 mM), the receptor population displayed monotonic binding parameters with a K/sub d/ for (/sup 3/H)-prazosin of 1.16 nM by Scatchard analysis. Competition curves against (/sup 3/H)-prazosin with the antagonists phentolamine and yohimbine revealed respective K/sub i/'s of .089 ..mu..M and 1.8 ..mu..M; curves with the agonists norepinephrine and isoproterenol yielded respective K/sub i/'s of 6.2..mu..M and 360 ..mu..M. Competition curves performed in the absence of guanyl nucleotide were complex demonstrating an apparent increase in affinity for agonists and an apparent decrease in affinity for antagonists. These curve shifts are consistent with the conversion of receptor to and from the guanyl nucleotide-sensitive state as a function of competing ligand concentration.},
doi = {},
url = {https://www.osti.gov/biblio/5904399}, journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 46:6,
place = {United States},
year = {Fri May 01 00:00:00 EDT 1987},
month = {Fri May 01 00:00:00 EDT 1987}
}

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