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(/sup 3/H)MK-801 labels a site on the N-methyl-D-aspartate receptor channel complex in rat brain membranes

Abstract

The potent noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist (/sup 3/H)MK-801 bound with nanomolar affinity to rat brain membranes in a reversible, saturable, and stereospecific manner. The affinity of (/sup 3/H)MK-801 was considerably higher in 5 mM Tris-HCl (pH 7.4) than in previous studies using Krebs-Henseleit buffer. (/sup 3/H)MK-801 labels a homogeneous population of sites in rat cerebral cortical membranes with KD of 6.3 nM and Bmax of 2.37 pmol/mg of protein. This binding was unevenly distributed among brain regions, with hippocampus greater than cortex greater than olfactory bulb = striatum greater than medulla-pons, and the cerebellum failing to show significant binding. Detailed pharmacological characterization indicated (/sup 3/H)MK-801 binding to a site which was competitively and potently inhibited by known noncompetitive NMDA receptor antagonists, such as phencyclidine, thienylcyclohexylpiperidine (TCP), ketamine, N-allylnormetazocine (SKF 10,047), cyclazocine, and etoxadrol, a specificity similar to sites labelled by (/sup 3/H)TCP. These sites were distinct from the high-affinity sites labelled by the sigma receptor ligand (+)-(/sup 3/H)SKF 10,047. (/sup 3/H)MK-801 binding was allosterically modulated by the endogenous NMDA receptor antagonist Mg2+ and by other active divalent cations. These data suggest that (/sup 3/H)MK-801 labels a high-affinity site on the NMDA receptor channel complex, distinct from the NMDA recognition  More>>
Publication Date:
Jan 01, 1988
Product Type:
Journal Article
Reference Number:
NLM-88-89611; EDB-88-060700
Resource Relation:
Journal Name: J. Neurochem.; (United Kingdom); Journal Volume: 50:1
Subject:
59 BASIC BIOLOGICAL SCIENCES; ANTICONVULSANTS; BIOCHEMICAL REACTION KINETICS; RECEPTORS; TISSUE DISTRIBUTION; AFFINITY; BRAIN; CELL MEMBRANES; RATS; TRACER TECHNIQUES; TRITIUM COMPOUNDS; ANIMALS; BODY; CELL CONSTITUENTS; CENTRAL NERVOUS SYSTEM; CENTRAL NERVOUS SYSTEM DEPRESSANTS; DISTRIBUTION; DRUGS; ISOTOPE APPLICATIONS; KINETICS; LABELLED COMPOUNDS; MAMMALS; MEMBRANE PROTEINS; MEMBRANES; NERVOUS SYSTEM; ORGANIC COMPOUNDS; ORGANS; PROTEINS; REACTION KINETICS; RODENTS; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques
OSTI ID:
5640520
Research Organizations:
Neuroscience Research Centre, Harlow, England
Country of Origin:
United Kingdom
Language:
English
Other Identifying Numbers:
Journal ID: CODEN: JONRA
Submitting Site:
NLM
Size:
Pages: 274-281
Announcement Date:
Mar 01, 1988

Citation Formats

Wong, E H, Knight, A R, and Woodruff, G N. (/sup 3/H)MK-801 labels a site on the N-methyl-D-aspartate receptor channel complex in rat brain membranes. United Kingdom: N. p., 1988. Web. doi:10.1111/j.1471-4159.1988.tb13260.x.
Wong, E H, Knight, A R, & Woodruff, G N. (/sup 3/H)MK-801 labels a site on the N-methyl-D-aspartate receptor channel complex in rat brain membranes. United Kingdom. https://doi.org/10.1111/j.1471-4159.1988.tb13260.x
Wong, E H, Knight, A R, and Woodruff, G N. 1988. "(/sup 3/H)MK-801 labels a site on the N-methyl-D-aspartate receptor channel complex in rat brain membranes." United Kingdom. https://doi.org/10.1111/j.1471-4159.1988.tb13260.x.
@misc{etde_5640520,
title = {(/sup 3/H)MK-801 labels a site on the N-methyl-D-aspartate receptor channel complex in rat brain membranes}
author = {Wong, E H, Knight, A R, and Woodruff, G N}
abstractNote = {The potent noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist (/sup 3/H)MK-801 bound with nanomolar affinity to rat brain membranes in a reversible, saturable, and stereospecific manner. The affinity of (/sup 3/H)MK-801 was considerably higher in 5 mM Tris-HCl (pH 7.4) than in previous studies using Krebs-Henseleit buffer. (/sup 3/H)MK-801 labels a homogeneous population of sites in rat cerebral cortical membranes with KD of 6.3 nM and Bmax of 2.37 pmol/mg of protein. This binding was unevenly distributed among brain regions, with hippocampus greater than cortex greater than olfactory bulb = striatum greater than medulla-pons, and the cerebellum failing to show significant binding. Detailed pharmacological characterization indicated (/sup 3/H)MK-801 binding to a site which was competitively and potently inhibited by known noncompetitive NMDA receptor antagonists, such as phencyclidine, thienylcyclohexylpiperidine (TCP), ketamine, N-allylnormetazocine (SKF 10,047), cyclazocine, and etoxadrol, a specificity similar to sites labelled by (/sup 3/H)TCP. These sites were distinct from the high-affinity sites labelled by the sigma receptor ligand (+)-(/sup 3/H)SKF 10,047. (/sup 3/H)MK-801 binding was allosterically modulated by the endogenous NMDA receptor antagonist Mg2+ and by other active divalent cations. These data suggest that (/sup 3/H)MK-801 labels a high-affinity site on the NMDA receptor channel complex, distinct from the NMDA recognition site, which is responsible for the blocking action of MK-801 and other noncompetitive NMDA receptor antagonists.}
doi = {10.1111/j.1471-4159.1988.tb13260.x}
journal = []
volume = {50:1}
journal type = {AC}
place = {United Kingdom}
year = {1988}
month = {Jan}
}