Tamulevicius P, Lax E R, Müller A, Schriefers H
Biochem J. 1982 Aug 15;206(2):279-86. doi: 10.1042/bj2060279.
A method for the determination of specific oestrogen-receptor binding sites in rat liver nuclei is described. Nuclear receptors showed a high affinity for oestradiol (Kd approximately 3 x 10(-9)M), a low capacity, and a distinct specificity for substances with known oestrogenic and anti-oestrogenic activity. No sex differences were seen in the concentrations of nuclear receptors from either vehicle- or ethynyloestradiol-pretreated rats. Only a limited number of binding sites could be extracted with 0.4 M-KCl. The remaining sites, which were solubilized by sonication and treatment with deoxyribonuclease I, sedimented at 3-4 S. Of four oestrogens tested (oestradiol, ethynyloestradiol, diethylstilboestrol, tri-p-anisylchloroethylene), ethynyloestradiol was the most effective translocation agent in vivo, nuclear uptake occurring at doses below 1 microgram/rat; changes in salt extractability of nuclear receptors occurred at doses lower than those required to achieve absolute increases in nuclear receptor concentrations.
本文描述了一种测定大鼠肝细胞核中特异性雌激素受体结合位点的方法。核受体对雌二醇表现出高亲和力（解离常数Kd约为3×10⁻⁹M）、低容量，并且对具有已知雌激素和抗雌激素活性的物质具有明显的特异性。在给予载体或乙炔雌二醇预处理的大鼠中，未观察到核受体浓度存在性别差异。仅用0.4M-KCl可提取出有限数量的结合位点。其余通过超声处理和脱氧核糖核酸酶I处理而溶解的位点，沉降系数为3 - 4S。在所测试的四种雌激素（雌二醇、乙炔雌二醇、己烯雌酚、三对甲氧基苯基氯乙烯）中，乙炔雌二醇是体内最有效的转运剂，在剂量低于1微克/大鼠时即可发生核摄取；核受体盐提取能力的变化发生的剂量低于实现核受体浓度绝对增加所需的剂量。
