Demura T, Driscoll W J, Strott C A
Section on Adrenal Cell Biology, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
Endocrinology. 1989 May;124(5):2200-7. doi: 10.1210/endo-124-5-2200.
Nuclei purified from the guinea pig adrenal cortex contain a specific progesterone-binding activity which, based on enzyme degradation studies, appears to be proteinaceous. Saturation analysis revealed a Kd of about 15 nM and a binding capacity of about 33 pmol/mg DNA. The activity of the nuclear binding protein was specific essentially for progestational steroids; the two most potent progesterone competitors were 5 alpha-pregnane-3,20-dione and medroxyprogesterone (17 alpha-hydroxy-6 alpha-methylprogesterone), while 17 beta-estradiol, testosterone, cortisol, and other related steroids were poor competitors. The adrenocortical nuclear progesterone-binding protein was present to an equal extent in both male and female guinea pigs. The adrenocortical nuclear progesterone-binding protein differed from the classical progesterone receptor in that 1) the affinity of the adrenocortical binding protein for progesterone is an order of magnitude lower; 2) the potent synthetic progestin R5020 binds less tightly to the adrenocortical progesterone-binding protein; 3) the adrenocortical progesterone-binding protein is not modulated by estrogenic activity; 4) the adrenocortical progesterone-binding protein is more stable at 37 C; 5) the adrenocortical nuclear progesterone-binding protein is not salt extractable; and 6) Western blot analysis has revealed that an antiprogesterone receptor monoclonal antibody, which recognizes the guinea pig uterine classical nuclear progesterone receptor, does not recognize the adrenocortical nuclear progesterone-binding protein. Thus, the guinea pig adrenocortical nucleus contains a type of progesterone-binding protein that appears to be clearly different from the classical progesterone receptor.
从豚鼠肾上腺皮质纯化得到的细胞核含有一种特异性的孕酮结合活性,基于酶降解研究,该活性似乎是蛋白质性质的。饱和分析显示解离常数(Kd)约为15 nM,结合容量约为33 pmol/mg DNA。核结合蛋白的活性基本上对孕甾类激素具有特异性;两种最有效的孕酮竞争者是5α-孕烷-3,20-二酮和甲羟孕酮(17α-羟基-6α-甲基孕酮),而17β-雌二醇、睾酮、皮质醇和其他相关类固醇是较弱的竞争者。肾上腺皮质核孕酮结合蛋白在雄性和雌性豚鼠中含量相等。肾上腺皮质核孕酮结合蛋白与经典孕酮受体的不同之处在于:1)肾上腺皮质结合蛋白对孕酮的亲和力低一个数量级;2)强效合成孕激素R5020与肾上腺皮质孕酮结合蛋白的结合较松散;3)肾上腺皮质孕酮结合蛋白不受雌激素活性调节;4)肾上腺皮质孕酮结合蛋白在37℃时更稳定;5)肾上腺皮质核孕酮结合蛋白不能用盐提取;6)蛋白质免疫印迹分析显示,一种识别豚鼠子宫经典核孕酮受体的抗孕酮受体单克隆抗体不能识别肾上腺皮质核孕酮结合蛋白。因此,豚鼠肾上腺皮质核含有一种似乎与经典孕酮受体明显不同的孕酮结合蛋白。