Application of radioreceptor assay to the purification of silver carp gonadotrophin
Application of radioreceptor assay to the purification of silver carp gonadotrophin
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放射性受体测定法在鲢鱼促性腺激素纯化中的应用
DOI：
10.2331/suisan.51.405
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发表时间：
1985
影响因子：
0.4
通讯作者：
S. Ishii
中科院分区：
文献类型：
--
作者：
Makito Kobayashi;K. Aida;I. Hanyu;S. Ishii
用放射受体分析法成功地检测出了银鱼促性腺激素糖蛋白，具有较高的灵敏度和精密度。通过乙醇提取、离子交换层析、层析聚焦、凝胶过滤等步骤进行纯化。用苦味产卵器生物测定法和放射受体测定法测定了牛蛙促性腺激素的活性，并对牛蛙的FSH和LH进行了鉴别。促性腺激素活性仅在酸性组分中检测到。在整个纯化过程中，所有三种活性总是以大约相同的比例在同一组分中检测到，并且不同组分之间没有分离促性腺激素活性。因此，尽管我们使用了针对青蛙FSH和LH的放射受体分析方法，但我们未能在银鱼脑下垂体中显示两种促性腺激素，如FSH和LH。这些结果支持一种观点，即鱼脑垂体中只含有一种促性腺激素。用凝胶过滤的方法，估计出鲤鱼促性腺激素的相对分子质量为38,000。最终制剂的等电聚焦显示几个组分的等电点不同(4.7、4.8和5.1)。
Radioreceptor assay was successfully used to detect glycoprotein gonadotropin of silver carp Hypophthalmichthys molitrix with high sensitivity and precision. Purification was carried out by ethanol extraction, ion exchange chromatography, chromatofocusing and gel filtration. The gonadotropic potency was measured by bitterling ovipositor bioassay and radioreceptor assays which were used for the discrimination of bullfrog FSH and LH. Gonadotropic activity was detected only in acidic fractions. Throughout the purification processes, all the three activities were always detected in the same fractions at about the same ratio and there was no separation of the gonadotropin activities among fractions. Thus, even though we employed the radioreceptor assay methods specific to each of frog FSH and LH, we failed to show two types of gonadotropins such as FSH and LH in the silver carp pituitary gland. These results favour an idea that fish pituitary contains a single type of gonadotropin.The molecular weight of the silver carp gonadotropin was estimated to be 38, 000 by gel filtra-tion. Isoelectrofocusing of the final preparation showed several components with different pIs (4.7, 4.8 and 5.1).