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

Journal Article · Tue Mar 06 00:00:00 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), Vol. 29:9; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

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