Ng T B, Hon W K, Idler D R
Biochem Cell Biol. 1987 Apr;65(4):386-91. doi: 10.1139/o87-048.
The pituitaries of vitellogenic sockeye salmon (Oncorhynchus nerka) were extracted with a mixture of acetone, water, and hydrochloric acid. The precipitate which formed upon the addition of a copious volume of acetone to the extract, designated acid acetone powder, was subjected to salt fractionation and desalting, followed by ion-exchange chromatography on CM-cellulose. An unadsorbed fraction (S-1) and four adsorbed fractions (S-2, S-3, S-4 and S-5) were obtained. Adrenocorticotropic activity was detected in the fractions by their ability to stimulate isolated rat adrenal decapsular cells to produce corticosterone and by their immunoreactivities in an adrenocorticotropin-specific radioimmunoassay. The steroidogenic activities of all fractions, except S-4, were blocked by corticotropin inhibiting peptide. Opiate activity was detected in the fractions by their ability to inhibit the binding of either [3H]naloxone or (D-ala2, D-leu5)-[3H]enkephalin to rat brain membranes. There was a discrepancy in the potencies of the five fractions in the two opiate radioreceptor assays, indicating the presence of opiate peptides with different affinities of binding to the micron- and delta-opiate receptors of the rat brain. There was a separation between adrenocorticotropic and opiate receptor binding activities, suggesting that the activities were due to separate molecular entities.
用丙酮、水和盐酸的混合物提取产卵期红大马哈鱼(Oncorhynchus nerka)的垂体。向提取物中加入大量丙酮后形成的沉淀物,称为酸丙酮粉,对其进行盐分级分离和脱盐处理,然后在CM - 纤维素上进行离子交换色谱分析。得到一个未吸附级分(S - 1)和四个吸附级分(S - 2、S - 3、S - 4和S - 5)。通过各组分刺激分离的大鼠肾上腺被膜细胞产生皮质酮的能力以及它们在促肾上腺皮质激素特异性放射免疫测定中的免疫反应性来检测各组分中的促肾上腺皮质激素活性。除S - 4外,所有组分的类固醇生成活性均被促肾上腺皮质激素抑制肽阻断。通过各组分抑制[3H]纳洛酮或(D - ala2,D - leu5) - [3H]脑啡肽与大鼠脑膜结合的能力来检测各组分中的阿片样物质活性。在两种阿片样物质放射受体测定中,五个组分的效力存在差异,表明存在与大鼠脑的μ - 和δ - 阿片样物质受体具有不同结合亲和力的阿片样肽。促肾上腺皮质激素和阿片样物质受体结合活性之间存在分离,这表明这些活性是由不同的分子实体引起的。