兔睾丸雄激素结合蛋白（ABP）的纯化与特性分析

Purification and characterization of rabbit testicular androgen binding protein (ABP).

作者信息

Weddington S C, Brandtzaeg P, Sletten K, Christensen T, Hansson V

出版信息

Curr Top Mol Endocrinol. 1975;2:433-51. doi: 10.1007/978-1-4613-4440-7_31.

PMID:1242082

Abstract

Testicular androgen binding protein (ABP) was purified from the epididymis of 1500 adult rabbits by the sequential use of ammonium sulphate precipitation, ion exchange chromatography on DEAE cellulose, gel filtration on Sephadex G-200, hydroxyl-apatite chromatography and preparative polyacrylamide gel electrophoresis. This procedure yielded a 1000-fold increase in specific activity compared to that of the 1500,000 x g supernatant, and the recovery of active ABP was about 3-5%. ABP is acid glycoprotein with a molecular weight of 65-68,000 daltons. Antisera to rabbit ABP raised in quinea pigs inhibit 3H-DHT binding to ABP as measured by SS-PAGE. When diluted rabbit serum containing TeBG is treated with the same dilutions of these antisera, identical binding inhibition curves are found. Thus, ABP and TeBG in rabbits appear to possess identical immunological determinants.

摘要

通过依次使用硫酸铵沉淀、DEAE纤维素离子交换色谱、Sephadex G - 200凝胶过滤、羟基磷灰石色谱和制备性聚丙烯酰胺凝胶电泳，从1500只成年兔子的附睾中纯化出睾丸雄激素结合蛋白（ABP）。与150,000×g上清液相比，该方法使比活性提高了1000倍，活性ABP的回收率约为3 - 5%。ABP是一种酸性糖蛋白，分子量为65 - 68,000道尔顿。用豚鼠制备的抗兔ABP抗血清，通过SS - PAGE检测可抑制3H - DHT与ABP的结合。当用相同稀释度的这些抗血清处理含有睾酮雌二醇结合球蛋白（TeBG）的稀释兔血清时，会得到相同的结合抑制曲线。因此，兔子体内的ABP和TeBG似乎具有相同的免疫决定簇。