Schreiber J R, Nakamura K, Erickson G F
Steroids. 1982 May;39(5):569-84. doi: 10.1016/0039-128x(82)90057-5.
A soluble, thermolabile protein with characteristics typical of glucocorticoid receptors has been identified in the ovaries of estrogen-stimulated hypophysectomized immature rats. After the incubation of 3H- dexamethasone with ovarian cytosol, fractionation on a Sephadex G-200 column reveals a peak of radioactivity which elutes at the void volume. This peak, which represents saturable 3H-dexamethasone binding, disappears following heating (4 degrees C X 15 min) or treatment of the cytosol with pronase. Scatchard analysis of the 3H-dexamethasone binding to cytosol shows it to be high affinity (Kd=5.1 nM) and saturable, with 327 fmol binding sites/mg cytosol protein. Binding site number rises linearly with increasing cytosol protein concentrations. The relative abilities of various steroids to inhibit 3H-dexamethasone binding are: triamcinolone acetonide greater than or equal to dexamethasone greater than cortisol = progesterone greater than dihydro-testosterone greater than estradiol. This binding protein sediments at 9 S on a sucrose gradient, has a mean Stokes radius of 105 A on gel exclusion chromatography, and has a calculated molecular weight of 388,000 daltons and a frictional ratio of 2.1. 3H-Dexamethasone is not metabolized and does not bind specifically to serum. We have identified a protein in the rat ovary with characteristics of a glucocorticoid receptor and propose that this protein may be responsible for mediating direct effects of glucocorticoids on the ovary.
在雌激素刺激的垂体切除未成熟大鼠的卵巢中，已鉴定出一种具有糖皮质激素受体典型特征的可溶性、热不稳定蛋白。用3H-地塞米松与卵巢胞质溶胶孵育后，在Sephadex G-200柱上进行分级分离，发现放射性峰在空体积处洗脱。这个代表可饱和的3H-地塞米松结合的峰，在加热（4℃×15分钟）或用链霉蛋白酶处理胞质溶胶后消失。对3H-地塞米松与胞质溶胶结合的Scatchard分析表明，它具有高亲和力（Kd = 5.1 nM）且可饱和，每毫克胞质溶胶蛋白有327 fmol结合位点。结合位点数随胞质溶胶蛋白浓度增加呈线性上升。各种类固醇抑制3H-地塞米松结合的相对能力为：曲安奈德≥地塞米松＞皮质醇 = 孕酮＞二氢睾酮＞雌二醇。这种结合蛋白在蔗糖梯度上沉降系数为9 S，在凝胶排阻色谱上平均斯托克斯半径为105 Å，计算分子量为388,000道尔顿，摩擦比为2.1。3H-地塞米松未被代谢，也不与血清特异性结合。我们在大鼠卵巢中鉴定出一种具有糖皮质激素受体特征的蛋白，并提出这种蛋白可能负责介导糖皮质激素对卵巢的直接作用。
