Purification of the opiate receptor from rat brain.
The opiate receptor was purified from a Triton-solubilized preparation of rat neural membranes by the use of affinity chromatography. The affinity gel was prepared by coupling 14-beta-bromoacetamidomorphine, a newly synthesized ligand, to omega-aminohexyl-Sepharose. After elution of the nonspecific proteins with 50 mM Tris (pH 7.5), the receptor proteins were eluted with 1 microM levorphanol or etorphine. NaDodSO4/polyacrylamide gel electrophoresis revealed three major proteins associated with the opiate receptor, having molecular weights of 43,000, 35,000, and 23,000. The purified receptor binds 10(-11) mol of dihydromorphine/per mg of protein, with a Kd of 3.8 X 10(-9) M. Other opiates, naloxone, and methionine-enkephalin, inhibit [3H]dihydromorphine binding in a manner similar to that observed with intact and solubilized neural membranes.
D000818:Animals; D001923:Brain Chemistry; D002462:Cell Membrane / Q000032:analysis; D002846:Chromatography, Affinity; D004090:Dihydromorphine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005048:Etorphine / Q000378:metabolism; D007981:Levorphanol / Q000378:metabolism; D009022:Morphine Derivatives / Q000378:metabolism; D051381:Rats; D011957:Receptors, Opioid / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
D009022:Morphine Derivatives; D011957:Receptors, Opioid; D007981:Levorphanol; D005048:Etorphine; C030452:14 beta-bromoacetamidomorphine; D004090:Dihydromorphine