N-methyl-D-aspartate/phencyclidine receptor complex of rat forebrain: purification and biochemical characterization.
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 67,000, 57,000, 46,000, 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.
Ikin
A F
AF
Department of Biochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel.
Kloog
Y
Y
Sokolovsky
M
M
D000818:Animals; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005971:Glutamates / Q000378:metabolism; D005998:Glycine / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D016202:N-Methylaspartate; D010622:Phencyclidine / Q000031:analogs & derivatives / Q000378:metabolism; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate; D017981:Receptors, Neurotransmitter / Q000302:isolation & purification / Q000378:metabolism*; D011957:Receptors, Opioid / Q000302:isolation & purification / Q000378:metabolism*; D017478:Receptors, Phencyclidine; D012995:Solubility
D005971:Glutamates; D016194:Receptors, N-Methyl-D-Aspartate; D017981:Receptors, Neurotransmitter; D011957:Receptors, Opioid; D017478:Receptors, Phencyclidine; D001224:Aspartic Acid; D016202:N-Methylaspartate; C012058:tenocyclidine; D010622:Phencyclidine; D005998:Glycine