Eisen L P, Harmon J M
Endocrinology. 1986 Oct;119(4):1419-26. doi: 10.1210/endo-119-4-1419.
Activation of the rat kidney mineralocorticoid receptor was investigated using diethylaminoethyl (DEAE)-cellulose, DNA-cellulose, and gel permeation chromatography. Specific labeling of the mineralocorticoid receptor was achieved by labeling with [3H]aldosterone in the presence of the pure glucocorticoid RU28362. The specificity of labeling was confirmed by the lack of immunoreactivity of [3H]aldosterone-labeled material with the monoclonal antiglucocorticoid receptor antibody BURG-1. The unactivated aldosterone-mineralocorticoid receptor complex did not bind to DNA-cellulose, was eluted from DEAE-cellulose at relatively high salt (195 mM KCl) concentration, and had an apparent Stokes radius when chromatographed on Sephacryl S300 of 6.3 nm. After activation, the aldosterone-mineralocorticoid receptor complex had increased affinity for DNA-cellulose and decreased affinity for DEAE-cellulose and appeared as a smaller complex when chromatographed on Sephacryl S300. These changes were blocked by sodium molybdate. Our results indicate that activation of the rat kidney mineralocorticoid receptor is analogous to activation of the glucocorticoid receptor and suggest that activation of the mineralocorticoid receptor involves dissociation of a multimeric receptor form.
使用二乙氨基乙基（DEAE）-纤维素、DNA-纤维素和凝胶渗透色谱法研究了大鼠肾脏盐皮质激素受体的激活情况。在纯糖皮质激素RU28362存在下，用[3H]醛固酮进行标记，实现了盐皮质激素受体的特异性标记。通过[3H]醛固酮标记的物质与单克隆抗糖皮质激素受体抗体BURG-1缺乏免疫反应性，证实了标记的特异性。未活化的醛固酮-盐皮质激素受体复合物不与DNA-纤维素结合，在相对高盐（195 mM KCl）浓度下从DEAE-纤维素上洗脱下来，在Sephacryl S300上进行色谱分析时，其表观斯托克斯半径为6.3 nm。激活后，醛固酮-盐皮质激素受体复合物对DNA-纤维素的亲和力增加，对DEAE-纤维素的亲和力降低，在Sephacryl S300上进行色谱分析时呈现为较小的复合物。这些变化被钼酸钠阻断。我们的结果表明，大鼠肾脏盐皮质激素受体的激活类似于糖皮质激素受体的激活，并表明盐皮质激素受体的激活涉及多聚体受体形式的解离。
