N-Methyl-D-aspartate/Phencyclidine Receptor Complex of Rat Forebrain: Purification and Biochemical Characterization

Annat F. Ikin, Yoel Kloog, Mordechai Sokolovsky

Research output: Contribution to journalArticlepeer-review

Abstract

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 3700-fold purification was achieved. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and dithiothreitol revealed four major bands of Mr 67000, 57 000, 46000, and 33 000. [3H]Azido-PCP was irreversibly incorporated into each of these bands after UV irradiation. The dissociation constant (Kd) of [1-(2-thienyl)cyclohexyl]piperidine ([3H]TCP) binding to the purified NMDA/PCP receptor was 120 nM. The maximum specific binding (Bmax) for [3H]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 [3H]TCP to the purified receptor was modulated by the NMDA receptor ligands glutamate, glycine, and NMDA.

Original languageEnglish
Pages (from-to)2290-2295
Number of pages6
JournalBiochemistry
Volume29
Issue number9
DOIs
StatePublished - 1 Mar 1990

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