Solubilization in high yield of opioid receptors retaining high-affinity delta, mu and kappa binding sites.
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.
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
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