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从兔子宫内膜分离和纯化细胞核,并在离体条件下观察雌二醇受体复合物与核结合作用的条件。核结合作用的适宜条件是在0℃和生理盐浓度,此时受体复合物和核表现稳定。胞液受体经25℃预温活化后能有效地与核进行结合作用,并增加与核的结合量。不同盐浓度对核结合作用有明显的影响。当KCl浓度逐步增加时,受体的结合量递减,这可能反映它们之间主要以离子键形式相互作用。在生理盐浓度条件下受体与核在0℃的结合作用表现饱和现象,随着加入受体量的增加显示两个坪区。Scatchard作图分析,表明核中存在多于一种的受体结合位点。一类数量较少(3,800位点/核),其结合亲和力较高,K_d值为3.4×10_(-10)M;另一类数量较多(48,400位点/核),其结合亲和力相对地较低,K_d值为1.49×10~(-9)M。
The nuclei were isolated and purified from the rabbit endometrium, and the conditions of estradiol receptor complex binding to the nucleus were observed under ex vivo conditions. Suitable conditions for nuclear binding are at 0 ° C and physiological salt concentrations, at which time the receptor complex and nucleus are stable. Activation of cytosolic receptors at 25 ℃ after pre-heating can effectively combine with the nucleus, and increase the binding capacity with the nucleus. Different salt concentrations have a significant effect on nuclear binding. As the KCl concentration gradually increases, the binding capacity of the receptors decreases, which probably reflects the predominantly ionic interactions between them. At physiological salt concentrations, the binding of the receptor to the nucleus saturates at 0 ° C, showing two plateaus as the amount of receptor added increases. Scatchard mapping analysis showed that there was more than one receptor binding site in the nucleus. One group with a small number (3,800 loci / nucleus) had a higher binding affinity with a K_d value of 3.4 × 10_ (-10) M and a larger number (48,400 loci / nucleus) with a higher binding affinity Lower, K_d value of 1.49 × 10 ~ (-9) M.