Banerjee P P, Bhattacharya S, Nath P
Department of Zoology, School of Life Science, Visva-Bharati University, West Bengal, India.
Gen Comp Endocrinol. 1989 Jan;73(1):118-28. doi: 10.1016/0016-6480(89)90062-2.
Gonadotropic hormone (GtH) from the pituitaries of two widely different Indian teleosts, a freshwater murrel and a carp, was purified by solvent fractionation, Sephadex G-100 gel filtration, affinity chromatography on concanavalin A-Sepharose (Con A-Sepharose), and immunoaffinity chromatography. Elution profile from gel filtration showed three peaks in both cases, peak I and II were clearly separated in murrel but not in carp. Peak II demonstrated strong GtH activity in both murrel and carp but this activity could also be detected in peak I and III of carp. TSH activity was restricted to peak I in murrel and was distributed in all three peaks in carp. Chromatography on Con A-Sepharose was useful in harvesting carp glycoprotein hormones, but gave no advantage in murrel. A pure homogenous GtH from murrel and carp could be obtained by using immunoaffinity chromatography. Polyacrylamide gel electrophoresis of murrel and carp GtH showed a single discrete band. Determination of molecular weight (MW) by Sephadex G-100 gel filtration indicated that murrel and carp GtH were 42,000 and 40,000 Da, respectively. SDS-polyacrylamide gel electrophoresis of murrel and carp GtH revealed two dissimilar subunits, alpha and beta. MW of murrel and carp alpha-subunits were 18,000 and 16,000 while those of beta were 27,000 and 26,000 Da, respectively. Comparison of electrophoretic patterns of murrel and carp GtH alpha and beta with ovine LH subunits reveal distinct teleostean GtH subunits.
从两种差异很大的印度硬骨鱼（一种淡水鲮鱼和一种鲤鱼）的垂体中提取的促性腺激素（GtH），通过溶剂分级分离、Sephadex G - 100凝胶过滤、伴刀豆球蛋白A - 琼脂糖（Con A - 琼脂糖）亲和层析和免疫亲和层析进行纯化。凝胶过滤的洗脱图谱在两种情况下均显示出三个峰，峰I和峰II在鲮鱼中明显分离，但在鲤鱼中未分离。峰II在鲮鱼和鲤鱼中均表现出较强的GtH活性，但在鲤鱼的峰I和峰III中也能检测到这种活性。促甲状腺激素（TSH）活性在鲮鱼中仅限于峰I，而在鲤鱼中分布于所有三个峰中。Con A - 琼脂糖层析有助于收获鲤鱼糖蛋白激素，但对鲮鱼没有优势。通过免疫亲和层析可获得来自鲮鱼和鲤鱼的纯的均一GtH。鲮鱼和鲤鱼GtH的聚丙烯酰胺凝胶电泳显示出一条单一的离散带。通过Sephadex G - 100凝胶过滤测定分子量（MW）表明，鲮鱼和鲤鱼GtH分别为42,000和40,000道尔顿。鲮鱼和鲤鱼GtH的SDS - 聚丙烯酰胺凝胶电泳显示出两种不同的亚基，α和β。鲮鱼和鲤鱼α亚基的MW分别为18,000和16,000，而β亚基的MW分别为27,000和26,000道尔顿。将鲮鱼和鲤鱼GtH的α和β亚基的电泳图谱与绵羊促黄体激素（LH）亚基进行比较，揭示了明显不同的硬骨鱼GtH亚基。
