N-methyl-D-aspartate/phencyclidine receptor complex of rat forebrain: Purification and biochemical characterization

Journal Article · Tue Mar 06 04:00:00 EST 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00461a012· OSTI ID:7118142

Ikin, A F; Kloog, Y; Sokolovsky, M ^[1]

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1. Tel Aviv Univ. (Israel)

The N-methyl-D-aspartate NMDA/phencyclidine (PCP) receptor from rat forebrain was solubilized with sodium cholate and purified by affinity chromatography on amino-PCP-agarose. A 3,700-fold purification was achieved. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and dithiothreitol revealed four major bands of M{sub r} 67,000, 57,000, 46,000, and 33,000. ({sup 3}H)Azido-PCP was irreversibly incorporated into each of these bands after UV irradiation. The dissociation constant (K{sub d}) of (1-(2-thienyl)cyclohexyl)piperidine (({sup 3}H)TCP) binding to the purified NMDA/PCP receptor was 120 nM. The maximum specific binding (B{sub max}) for ({sup 3}H)TCP binding was 3.3 nmol/mg of protein. The pharmacological profile of the purified receptor complex was similar to that of the membranal and soluble receptors. The binding of ({sup 3}H)TCP to the purified receptor was modulated by the NMDA receptor ligands glutamate, glycine, and NMDA.

• OSTI ID: 7118142
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 29:9; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
AMINO ACIDS
ANIMALS
ASPARTIC ACID
AZIDO COMPOUNDS
AZINES
BODY
BRAIN
CARBOXYLIC ACIDS
CENTRAL NERVOUS SYSTEM
ELECTROPHORESIS
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
MAMMALS
MEMBRANE PROTEINS
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PHARMACOLOGY
PIPERIDINES
PROTEINS
PURIFICATION
PYRIDINES
RADIORECEPTOR ASSAY
RATS
RECEPTORS
RODENTS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES