Romanova N A
Probl Endokrinol (Mosk). 1980 Mar-Apr;26(2):60-6.
A 300-fold purification of glucocorticosteroid-receptor complexes (GRC) of the rat liver with the 10--25% yield was conducted by combining the methods of binding with DNA-cellulose and salting out with ammonium sulfate at 30% saturation. An important feature of this method was a preponderant removal of protein admixtures capable of dissociation with DNA. In comparison of column chromatography and batch-procedure the latter proved to possess a number of advantages. Some properties of partially purified GRC were also studied. As revealed, the purified GRC which rapidly decomposed at 2 degrees C, could be partially stabilized by addition of inert protein, glycerine, SH-reagents and an excess of dexamethazone. Under such conditions about 13% of the initial amount of purified GRC was preserved after a week of storage at 2 degrees C. Determination of purified GRC content in the preparation by sorption on dextran coated carbon gave sharply diminished results; as to the method of oxyapatite sorption--it proved to be adequate. In comparing the GRC properties without and after the purification demonstrated that sedimentation and other properties of these preparations largely coincided. GRC-acceptor capacity of one- and two-chain DNA depended to a different degree on cytoplasmic admixtures in the GRC preparation removed in its purification.
通过结合与DNA纤维素结合的方法和用30%饱和度的硫酸铵盐析的方法，对大鼠肝脏的糖皮质激素受体复合物（GRC）进行了300倍的纯化，产率为10%-25%。该方法的一个重要特点是能大量去除可与DNA解离的蛋白质杂质。与柱色谱法和分批法相比，后者具有许多优点。还研究了部分纯化的GRC的一些性质。结果表明，纯化后的GRC在2℃下会迅速分解，通过添加惰性蛋白质、甘油、巯基试剂和过量的地塞米松可使其部分稳定。在这种条件下，纯化后的GRC在2℃下储存一周后，约13%的初始量得以保留。通过葡聚糖包被碳吸附法测定制剂中纯化后的GRC含量，结果明显降低；至于羟基磷灰石吸附法，结果证明是合适的。比较纯化前后GRC的性质表明，这些制剂的沉降和其他性质在很大程度上是一致的。单链和双链DNA的GRC受体能力在不同程度上取决于纯化过程中去除的GRC制剂中的细胞质杂质。
