Golsteyn E J, Lefebvre Y A
Biochim Biophys Acta. 1985 Feb 14;812(3):679-87. doi: 10.1016/0005-2736(85)90261-5.
Purified nuclear envelopes have been isolated from an androgen-responsive and two androgen-unresponsive cell lines of the Shionogi mouse mammary carcinoma. The binding of dihydrotestosterone to nuclear envelope fractions isolated from the three variant cell lines is correlated with the androgen-responsiveness of the cell line. Nuclear envelopes prepared from the two androgen-unresponsive cell lines did not bind dihydrotestosterone specifically following incubation with radioactive dihydrotestosterone from 2.5 to 45.0 nM at 20 degrees C for 18 h. Under the same binding conditions, nuclear envelopes prepared from the androgen-responsive cell line demonstrated saturable, specific binding of dihydrotestosterone. Scatchard analysis revealed a class of binding sites with an apparent Kd of 14.2 nM and a maximum binding capacity of 28.7 fmol/mg protein. Proteinase and heat treatments resulted in the complete loss of androgen-binding activity, whereas DNAase treatment resulted in the loss of 38% of the binding activity. The binding sites were specific for dihydrotestosterone. Testosterone was only a weak competitor and estradiol did not compete. Extraction with concentrations of KCl up to 1.0 M did not result in loss of androgen binding.
已从日本盐野义小鼠乳腺癌的一个雄激素反应性细胞系和两个雄激素无反应性细胞系中分离出纯化的核膜。从这三个变异细胞系中分离得到的核膜组分与二氢睾酮的结合情况与细胞系的雄激素反应性相关。在20℃下，将来自两个雄激素无反应性细胞系制备的核膜与2.5至45.0 nM的放射性二氢睾酮孵育18小时后，未检测到二氢睾酮的特异性结合。在相同的结合条件下，来自雄激素反应性细胞系制备的核膜表现出二氢睾酮的可饱和特异性结合。Scatchard分析显示存在一类结合位点，其表观解离常数（Kd）为14.2 nM，最大结合容量为28.7 fmol/mg蛋白质。蛋白酶和热处理导致雄激素结合活性完全丧失，而脱氧核糖核酸酶处理导致38%的结合活性丧失。这些结合位点对二氢睾酮具有特异性。睾酮只是一个弱竞争者，而雌二醇不参与竞争。用浓度高达1.0 M的氯化钾提取不会导致雄激素结合丧失。
