Ito Y, Makimura M, Murakoshi Y
Neuropeptides. 1984 Dec;5(1-3):193-6. doi: 10.1016/0143-4179(84)90060-x.
In brain membrane preparations, 3H-naloxone binding to bullfrog brain gave the highest values versus other species such as carp, chicken, rat and mouse. Scatchard analysis revealed that the Bmax value of bullfrog brain membranes was three fold greater than that of rat brain membranes. Opioid receptors were solubilized from bullfrog brain membranes using digitonin or 3-(cholamidopropyl)-dimethylammonio-1-propane sulfonate (CHAPS) as the detergent. The properties of solubilized opioid receptors were essentially the same as membrane bound receptors with regard to sensitivity to various treatments. The order of potency for a series of agonists in displacing 3H-naloxone binding in both membrane and solubilized preparations was bremazocine greater than pentazocine greater than morphine greater than delta-receptor peptide. Partial purification of opioid receptors was performed using a fast protein liquid chromatography system. Specific binding activity of 3H-naloxone was increased 10-15 fold within 20 min using the Mono Q column.
在脑膜制剂中，与鲤鱼、鸡、大鼠和小鼠等其他物种相比，3H-纳洛酮与牛蛙脑的结合值最高。Scatchard分析表明，牛蛙脑膜的Bmax值比大鼠脑膜的大三倍。使用洋地黄皂苷或3-(胆酰胺丙基)-二甲基氨基-1-丙烷磺酸盐（CHAPS）作为去污剂，从牛蛙脑膜中溶解阿片受体。就对各种处理的敏感性而言，溶解的阿片受体的性质与膜结合受体基本相同。在膜和溶解制剂中，一系列激动剂取代3H-纳洛酮结合的效力顺序为布瑞马唑辛大于喷他佐辛大于吗啡大于δ-受体肽。使用快速蛋白质液相色谱系统对阿片受体进行了部分纯化。使用Mono Q柱，3H-纳洛酮的特异性结合活性在20分钟内增加了10至15倍。
