title: Solubilization in high yield of opioid receptors retaining high-affinity delta, mu and kappa binding sites.
abstract: The binding sites for opiates (agonist and antagonist) and opioid peptides can be solubilized from rat brain membranes with digitonin in the presence of Mg2+ (10 mM). High affinity and high capacity binding to the soluble delta, mu, and kappa receptors is obtainable when the membranes are treated in Mg2+ (30 degrees C, 60 min) prior to solubilization. The yields of solubilized binding sites extracted with digitonin, 40-90%, are higher than those obtained from Mg2+-pretreated membranes with other detergents commonly used for receptor solubilization. The stability of the digitonin-soluble opioid receptor at room temperature makes it useful for purification and characterization.
mesh_terms: D000818:Animals; D001665:Binding Sites; D001923:Brain Chemistry; D002462:Cell Membrane / Q000032:analysis; D011232:Chemical Precipitation; D003902:Detergents; D004072:Digitonin; D004355:Drug Stability; D004399:Dynorphins; D004723:Endorphins / Q000378:metabolism; D004743:Enkephalin, Leucine / Q000031:analogs & derivatives / Q000378:metabolism; D016308:Enkephalin, Leucine-2-Alanine; D005048:Etorphine / Q000378:metabolism; D008274:Magnesium; D010446:Peptide Fragments / Q000378:metabolism; D011092:Polyethylene Glycols; D051381:Rats; D011957:Receptors, Opioid / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
chemical_list: D003902:Detergents; D004723:Endorphins; D010446:Peptide Fragments; D011957:Receptors, Opioid; D011092:Polyethylene Glycols; D005048:Etorphine; D004743:Enkephalin, Leucine; D016308:Enkephalin, Leucine-2-Alanine; D004399:Dynorphins; D008274:Magnesium; D004072:Digitonin