Pousette A, Snochowski M, Bression D, Högberg B, Gustafsson J A
Biochim Biophys Acta. 1979 Jan 18;582(2):358-67. doi: 10.1016/0304-4165(79)90397-0.
3H-labelled R 1881 (methyltrienolone) was bound with high affinity (Kd, 2 . 10(-9) M) and low capacity (11--15 fmol/mg protein)in prostate cytosol from rats castrated 18 h earlier. The binding was inhibited by 5alpha-dihydrotestosterone, testosterone, cyproterone acetate and LEO 1727, but not by estradiol-17beta, R 5020 or corticosterone. The 3H-labelled R 1881-receptor complex was excluded from a Sephadex G=200 column, had a sedimentation coefficient of 3.5 S, was eluted from a DEAE-cellulose column at 0.2 M KCl concentration and precipitated between 20--40% of saturation of ammonium sulfate. 20% of specifically bound 3H-labelled R 1881 was retained by DNA-cellulose at low ionic strength. The fraction which was not bound to DNA-cellulose did not bind either, after rechromatography, not even following heating at 20 degrees C for 20 min or following fractionation with ammonium sulphate. In summary, the results indicate that the 3H-labelled R 1881-receptor complex in rat prostate cytosol has characteristics similar to those of the 5alpha-[3H]dihydrotestosterone-receptor complex.
3H标记的R 1881(甲基三烯醇酮)与18小时前阉割的大鼠前列腺胞质溶胶具有高亲和力(解离常数Kd为2×10⁻⁹ M)和低容量(11 - 15 fmol/mg蛋白质)结合。5α-二氢睾酮、睾酮、醋酸环丙孕酮和LEO 1727可抑制这种结合,但17β-雌二醇、R 5020或皮质酮则不能。3H标记的R 1881 - 受体复合物被排阻于葡聚糖G - 200柱外,沉降系数为3.5 S,在0.2 M KCl浓度下从DEAE - 纤维素柱上洗脱,在硫酸铵饱和度20% - 40%之间沉淀。在低离子强度下,20%特异性结合的3H标记的R 1881被DNA - 纤维素保留。未与DNA - 纤维素结合的部分,再经色谱分离后,即使在20℃加热20分钟或用硫酸铵分级分离后也不再结合。总之,结果表明大鼠前列腺胞质溶胶中3H标记的R 1881 - 受体复合物具有与5α - [3H]二氢睾酮 - 受体复合物相似的特性。