Reel J R, Shih Y
Acta Endocrinol (Copenh). 1975 Oct;80(2):344-54. doi: 10.1530/acta.0.0800344.
In order to characterize the uterine progesterone-binding proteins of oestrogen-primed and unprimed, ovariectomized immature and adult golden hamsters, cytosols were incubated with [3H]progesterone and/or other steroids and analyzed by sucrose-glycerol density gradient ultracentrifugation. Progesterone-binding components with sedimentation coefficients of 7S and 4.5S were found in the uterine cytosols, but not in the cytosols from the hypothalamus, pituitary, diaphragm, or small intestine. Oestradiol-17beta markedly elevated the level of 7S uterine receptor and this increase appeared to be due to new receptor synthesis, since actinomycin D and cycloheximide blocked this response. Fifty to 100 mug of oestradiol-17beta per kg body weight was found to promote a maximum increase in the 7S macromolecular component. The 7S receptor showed a tendency to saturate at 1 X 10(-7) M [1,2-3H]progesterone, indicating limited capacity. At a molar ratio of 100:1, unlabeled progesterone competed effectively for 7S and 4,5S [3H]progesterone binding, whereas 5alpha-pregnane-3,20-dione, oestradiol-17beta and testosterone did not. Moreover, [1,2-3H]cortisol and [1,2-3H]corticosterone did not bind to the 7S receptor, implying steroid specificity. CI-628, a non-steroid oestrogen antagonist, completely prevented [6,7-3H]oestradiol-17beta binding to its 8.5S uterine cytosol receptor, but was entirely without effect on 7S and 4.5S [3H]progesterone binding. Pronase, but not DNase or RNase, abolished 7S and 4.5S progesterone binding, suggesting that the binders are at least in part protein. Protamine sulphate and p-hydroxymercuribenzoate also obliterated 7S binding, implying that this receptor may be an acidic protein which contains sulfhydryl groups that are necessary for progesterone binding.
为了表征雌激素预处理和未预处理的去卵巢未成熟及成年金黄仓鼠子宫中的孕酮结合蛋白,将细胞溶质与[3H]孕酮和/或其他类固醇一起孵育,并通过蔗糖 - 甘油密度梯度超速离心进行分析。在子宫细胞溶质中发现了沉降系数为7S和4.5S的孕酮结合成分,但在下丘脑、垂体、膈肌或小肠的细胞溶质中未发现。雌二醇 - 17β显著提高了7S子宫受体的水平,这种增加似乎是由于新受体的合成,因为放线菌素D和环己酰亚胺阻断了这种反应。发现每千克体重50至100微克的雌二醇 - 17β可促进7S大分子成分的最大增加。7S受体在1×10(-7)M [1,2 - 3H]孕酮时显示出饱和趋势,表明结合能力有限。在100:1的摩尔比下,未标记的孕酮有效地竞争7S和4.5S [3H]孕酮结合,而5α - 孕烷 - 3,20 - 二酮、雌二醇 - 17β和睾酮则不能。此外,[1,2 - 3H]皮质醇和[1,2 - 3H]皮质酮不与7S受体结合,这意味着类固醇特异性。非甾体雌激素拮抗剂CI - 628完全阻止了[6,7 - 3H]雌二醇 - 17β与其8.5S子宫细胞溶质受体的结合,但对7S和4.5S [3H]孕酮结合完全没有影响。链霉蛋白酶而非脱氧核糖核酸酶或核糖核酸酶消除了7S和4.