Subunit structure and purification of opioid receptors.
Considerable evidence indicates the existence of multiple types of opioid receptors. The three major types have been named mu, delta and kappa. The earlier evidence was based on pharmacological as well as membrane binding experiments. This paper will emphasize more recent studies using solubilized opioid binding sites. Several laboratories, including our own, have succeeded in separating kappa receptors from other types. A similar separation of mu from delta receptors has not yet been achieved. By crosslinking experiments with 125I- human beta-endorphin we have been able to provide strong evidence for differences in molecular size between the major binding components of mu (65K) and delta (53K) receptors. It is not yet established whether the difference resides in the protein or carbohydrate portion of these glycoproteins. These results suggest that the three major types of opioid receptors represent distinct molecular entities. An active opioid binding protein solubilized from bovine striatal membranes has been purified to apparent homogeneity. The major purification steps involve affinity chromatography and lectin chromatography on immobilized wheat germ agglutinin. The purified material gave a single band of molecular weight 65K Da on SDS-PAGE. Its specific activity for opioid binding was ca. 13,000 pmol/mg protein and its properties are those of a component of the mu receptor.
D000818:Animals; D002417:Cattle; D003342:Corpus Striatum / Q000032:analysis; D004723:Endorphins / Q000378:metabolism; D004745:Enkephalins / Q000378:metabolism; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism; D049673:History, 20th Century; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D011487:Protein Conformation; D011957:Receptors, Opioid / Q000145:classification* / Q000266:history / Q000302:isolation & purification / Q000378:metabolism; D017465:Receptors, Opioid, delta; D017473:Receptors, Opioid, kappa; D017450:Receptors, Opioid, mu; D014909:Wheat Germ Agglutinins / Q000378:metabolism; D001615:beta-Endorphin
D004723:Endorphins; D004745:Enkephalins; D006023:Glycoproteins; D008565:Membrane Proteins; D011957:Receptors, Opioid; D017465:Receptors, Opioid, delta; D017473:Receptors, Opioid, kappa; D017450:Receptors, Opioid, mu; D014909:Wheat Germ Agglutinins; D001615:beta-Endorphin