Murakami N, Moudgil V K
Biochim Biophys Acta. 1981 Sep 4;676(3):386-94. doi: 10.1016/0304-4165(81)90175-6.
Glucocorticoid-receptor complex from rat liver cytosol, activated by warming at 23 degrees C or fractionation with (NH4)2SO4, was adsorbed over DNA-cellulose. This DNA-cellulose-bound [3H]triamcinolone acetonide-receptor complex was extracted in a dose-dependent manner by incubation with different concentrations of sodium tungstate. A 50% recovery of receptor was achieved with 5 mM sodium tungstate. Almost the entire glucocorticoid-receptor complex bound to DNA-cellulose could be extracted with 20 mM sodium tungstate. The [3H]triamcinolone acetonide released from DNA-cellulose following tungstate and molybdate treatment was found to be associated with a macromolecule, as seen by analysis on a Sephadex G-75 column. The glucocorticoid-receptor complex extracted by both the compounds sedimented as a 4 S entity of 5-20% sucrose gradients under low- and high-salt conditions. Addition of tungstate or molybdate to the preparations containing activated receptor had no effect on the sedimentation rate of receptor. However, addition of tungstate to non-activated receptor preparation caused aggregates of larger size. The tungstate-extracted glucocorticoid-receptor complex failed to rebind to DNA-cellulose even after extensive dialysis, whereas receptor in molybdate-extract retained its DNA-cellulose binding capacity.
通过在23℃加热或用硫酸铵分级分离激活的大鼠肝脏细胞质中的糖皮质激素受体复合物，被吸附在DNA纤维素上。这种结合在DNA纤维素上的[³H]曲安奈德受体复合物通过与不同浓度的钨酸钠孵育以剂量依赖的方式被提取出来。用5 mM钨酸钠可实现50%的受体回收率。用20 mM钨酸钠几乎可以提取出结合在DNA纤维素上的全部糖皮质激素受体复合物。经葡聚糖凝胶G-75柱分析发现，钨酸盐和钼酸盐处理后从DNA纤维素上释放的[³H]曲安奈德与一种大分子相关。在低盐和高盐条件下，这两种化合物提取的糖皮质激素受体复合物在5 - 20%蔗糖梯度中以4S实体形式沉降。向含有激活受体的制剂中添加钨酸盐或钼酸盐对受体的沉降速率没有影响。然而，向未激活的受体制剂中添加钨酸盐会导致形成更大尺寸的聚集体。经钨酸盐提取的糖皮质激素受体复合物即使经过广泛透析也无法重新结合到DNA纤维素上，而钼酸盐提取物中的受体保留了其与DNA纤维素的结合能力。
