Bidlack J M, Abood L G, Munemitsu S M, Archer S, Gala D, Kreilick R W
Adv Biochem Psychopharmacol. 1982;33:301-9.
Affinity labeling of the opiate receptor has been performed on neural membranes from rat brain utilizing[125I]-14-bromoacetamidomorphine, an opiate agonist, and [125I]-14-chloracetylmorphine, an antagonist. With the use of SDS gel electrophoresis it could be shown that the agonist labeled three proteins with molecular weights of 43,000, 35,000 and 23,000, whereas the antagonist only labeled the 23,000 component. The preferential labeling of the 23,000 protein by the antagonist suggests that this component may be a primary recognition site for opiate antagonists. Calcium was stimulatory to the affinity labeling of all three proteins while sodium was inhibitory. With the use of affinity columns prepared by conjugating either ligand to omega-aminohexyl Sepharose, a receptor complex was obtained consisting all three proteins. Stereospecific opiate binding was demonstrable in the complex prepared from either column.
利用阿片类激动剂[125I]-14-溴乙酰吗啡和拮抗剂[125I]-14-氯乙酰吗啡，对大鼠脑的神经膜进行了阿片受体的亲和标记。通过十二烷基硫酸钠凝胶电泳可以表明，激动剂标记了三种分子量分别为43000、35000和23000的蛋白质，而拮抗剂只标记了分子量为23000的成分。拮抗剂对分子量为23000的蛋白质的优先标记表明，该成分可能是阿片类拮抗剂的主要识别位点。钙对所有三种蛋白质的亲和标记有刺激作用，而钠则有抑制作用。通过将两种配体与ω-氨基己基琼脂糖偶联制备亲和柱，获得了由所有三种蛋白质组成的受体复合物。从任一柱制备的复合物中均可证明存在立体特异性阿片结合。
