Solubilization of phencyclidine receptors from rat cerebral cortex in an active ligand binding state.
A phencyclidine (PCP) receptor binding site has been solubilized in an active ligand-binding state from rat cerebral cortical membranes with sodium deoxycholate. Optimal receptor solubilization occurs at a detergent/protein ratio of 0.5 (w/w); for 5 mg protein/ml solubilized with 0.25% sodium deoxycholate, about 60% of the protein and 25% of the receptor is solubilized. Specific binding of either [3H]-N-[1-(2-thienyl)cyclohexyl]piperidine ([3H]TCP) or [3H]MK-801 is measurable by filtration through Sephadex G-50 columns or glass fiber filters; more than 60% of the binding activity is stable after 48 h at 4 degrees C. In the presence of detergent, [3H]TCP binding exhibits a Kd of 250 nM, a Bmax of 0.56 pmol/mg protein, and a pharmacological profile consistent with that of the membrane-bound PCP receptor, although most drugs bind with affinities 2 to 8 fold lower than in membranes. Upon reduction of detergent concentration, binding parameters approximate those for the membrane-bound receptor ([3H]TCP binding: Kd = 48 nM, Bmax = 1.13 pmol/mg protein).
McVittie
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National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, MD 20892.
Sibley
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D000818:Animals; D001681:Biological Assay; D002540:Cerebral Cortex / Q000378:metabolism*; D002850:Chromatography, Gel; D003840:Deoxycholic Acid / Q000378:metabolism; D003902:Detergents; D008024:Ligands / Q000378:metabolism*; D051381:Rats; D017981:Receptors, Neurotransmitter / Q000378:metabolism*; D017478:Receptors, Phencyclidine; D012995:Solubility; D013696:Temperature; D013997:Time Factors
D003902:Detergents; D008024:Ligands; D017981:Receptors, Neurotransmitter; D017478:Receptors, Phencyclidine; D003840:Deoxycholic Acid