Stabilization of 8S progesterone receptor from human prostate in the presence of molybdate ion.
Stabilization of 8S progesterone receptor from human prostate in the presence of molybdate ion.
复制标题
在钼酸根离子存在下稳定人前列腺 8S 孕酮受体。
作者：
C. Bevins;N. Bashirelahi
在10 mM钼酸根离子存在下，我们能够在人的前列腺胞浆液中检测到[6，7-3H]-17，21-dimethyl-19-nor-4，9-pregnadiene-3，20-dione-([3H]R5020)结合部分的出现，该部分在甘油密度梯度中以大约8s的速度沉淀。特异性结合的[~3H]-R5020可被黄体酮、替拉西松酮和R5020取代，但不能被皮质醇、双氢睾酮、17β-雌二醇或己烯雌酚取代。在钼酸根离子存在下，[~3H]R5020的结合在7×10~(-10)M处达到饱和(n=fmol/mg蛋白质)。确定了增强[~3H]-R5020特异性结合的最佳钼酸根离子浓度为7~20 mM。在没有添加类固醇的情况下，钼酸根离子在0℃预孵育16小时也能有效地稳定8S R5020结合部分。
In the presence of 10 mM molybdate ion, we are able to detect the appearance of a [6,7-3H]-17,21-dimethyl-19-nor-4,9-pregnadiene-3,20-dione-([3H]R5020) binding moiety in human prostatic cytosol which sedimented at approximately 8S in a glycerol density gradient. The specifically bound [3H]-R5020 was displaced by progesterone, tiramcinolone acetonide, and R5020 but not by cortisol, dihydrotestosterone, 17 beta-estradiol, or diethylstilbestrol. Specific binding of [3H]R5020 in the presence of molybdate ion was shown to saturate at 7 x 10(-10) M (n = 64 fmol/mg of protein). The optimum concentration of molybdate ion for enhancing specific binding of [3H]-R5020 was determined to be between 7 and 20 mM. Molybdate ion was also effective in stabilizing the 8S R5020-binding moiety in a preincubation for 16 hr at 0 degrees in the absence of added steroid.