Solubilization and characterization of sigma-receptors from guinea pig brain membranes.

Details

Serval ID
serval:BIB_0DD46C089399
Type
Article: article from journal or magazin.
Collection
Publications
Title
Solubilization and characterization of sigma-receptors from guinea pig brain membranes.
Journal
Journal of neurochemistry
Author(s)
Kavanaugh M.P., Parker J., Bobker D.H., Keana J.F., Weber E.
ISSN
0022-3042 (Print)
ISSN-L
0022-3042
Publication state
Published
Issued date
11/1989
Peer-reviewed
Oui
Volume
53
Number
5
Pages
1575-1580
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
The sigma-receptor, a distinct binding site in brain tissue that may mediate some of the psychotomimetic properties of benzomorphan opiates and phencyclidine, has been solubilized using the ionic detergent sodium cholate. Binding assays were performed with the solubilized receptor using vacuum filtration over polyethyleneimine-treated glass fiber filters. The pharmacological specificity of the solubilized binding site for sigma-receptor ligands is nearly identical to the membrane-bound form of the receptor, with the order of potencies for displacement of the selective sigma-ligand [3H]di-o-tolylguanidine ([3H]DTG) closely correlated. The stereoselectivity for (+)-benzomorphan opiate enantiomers was retained by the solubilized receptor. The soluble receptor retained high affinity for binding of [3H]DTG (KD = 28 +/- 0.5 nM) and (+)-[3H]3-(3-hydroxyphenyl)-N-(1-propyl)piperidine [(+)-[3H]3-PPP] (KD = 36 +/- 2 nM). Photoaffinity labeling of the solubilized receptor by [3H]p-azido-DTG, a sigma-selective photoaffinity label, resulted in labeling of a 29-kilodalton polypeptide identical in size to that labeled in intact membranes. Estimation of the Stokes radius of the [3H]DTG binding site was obtained by Sepharose CL-6B chromatography in the presence of 20 mM cholate and calculated to be 8.7 nm. This value was identical to the molecular size found for the binding sites of the sigma-selective ligands (+)-[3H]3-PPP and (+)-[3H]SKF-10,047, supporting the hypothesis that all three ligands bind to the same macromolecular complex.
Keywords
Affinity Labels, Animals, Binding Sites, Brain/metabolism, Cell Membrane/metabolism, Chemical Phenomena, Chemistry, Chromatography, Gel, Guanidines/metabolism, Guinea Pigs, Receptors, Opioid/metabolism, Receptors, sigma, Solubility
Pubmed
Web of science
Create date
19/11/2007 9:28
Last modification date
14/04/2023 10:48
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