Pfahl M, Sandros T, Bourgeois S
Mol Cell Endocrinol. 1978 Apr;10(2):175-91. doi: 10.1016/0303-7207(78)90124-7.
Procedures have been developed which provide simple means of determining binding constants of steroid receptors for glucocorticoids in mouse lymphoid cell lines and of characterizing the interaction of the steroid--receptor complex with the nucleus. An average of 70% of the steroid--receptor complexes is found associated with the nuclear fraction in three investigated cell lines, whereas 30% of the steroid--receptor complexes is found in the cytosol fraction. This distribution of the steroid-receptor complex within the cell is independent of whether steroid uptake of the cells is performed at low or at high steroid concentration. Part of the binding of the steroid receptor to the nuclear fraction is sensitive to high ionic strength and to high pH. A larger fraction of the steroid--receptor complex binding to the nuclear fraction is insensitive to high ionic strength and pH when the steroid uptake is performed at low steroid concentrations than when performed at high steroid concentrations. Steroid--receptor complex is released from the nuclear fraction by DNAase treatment but not by RNAase treatment. The possible correlation between the sensitivity to ionic strength and pH and the specificity of the binding is discussed.
已开发出一些方法，这些方法提供了简单的手段来测定小鼠淋巴细胞系中糖皮质激素的类固醇受体结合常数，并表征类固醇 - 受体复合物与细胞核的相互作用。在所研究的三种细胞系中，平均70%的类固醇 - 受体复合物与细胞核部分相关联，而30%的类固醇 - 受体复合物存在于胞质溶胶部分。细胞内类固醇 - 受体复合物的这种分布与细胞是在低类固醇浓度还是高类固醇浓度下摄取类固醇无关。类固醇受体与细胞核部分的部分结合对高离子强度和高pH敏感。当在低类固醇浓度下进行类固醇摄取时，与在高类固醇浓度下进行摄取相比，结合到细胞核部分的类固醇 - 受体复合物的更大比例对高离子强度和pH不敏感。类固醇 - 受体复合物通过DNA酶处理从细胞核部分释放，但不通过RNA酶处理释放。讨论了对离子强度和pH的敏感性与结合特异性之间的可能相关性。
