Logeat F, Le Cunff M, Pamphile R, Milgrom E
Biochem Biophys Res Commun. 1985 Aug 30;131(1):421-7. doi: 10.1016/0006-291x(85)91819-4.
The solubilized ("cytosolic") receptor present in the rabbit uterus in the absence of hormone and the chromatin-bound ("nuclear") receptor obtained after injection of a progestin were compared. Crude cellular extracts were analyzed by immunoblotting and receptors were purified by immunoaffinity chromatography. With both methods it was observed that the electrophoretic mobility of the "nuclear" receptor was slower than that of the "cytosolic" receptor. This difference in mobility appeared to be due to the existence of variably phosphorylated forms of receptor. The phosphorylation reaction was examined in uterine slices. In the absence of hormone the cytosolic receptor was phosphorylated. When hormone was added the phosphorylation of receptor was markedly enhanced and the electrophoretic mobility of the "nuclear" receptor was decreased. These experiments thus show that the receptor in its "cytosolic" form is a phosphoprotein. Under the effect of the hormone the receptor is further phosphorylated on some supplementary site(s). This polyphosphoprotein is the chromatin-bound, putatively active, form of the receptor. In this respect the intracellular progesterone receptor is similar to various membrane receptors for hormones and growth factors which are phosphorylated upon binding of their ligand.
对兔子宫中在无激素情况下存在的可溶性(“胞质”)受体与注射孕激素后获得的染色质结合(“核”)受体进行了比较。通过免疫印迹分析粗细胞提取物,并通过免疫亲和层析纯化受体。使用这两种方法均观察到“核”受体的电泳迁移率比“胞质”受体慢。迁移率的这种差异似乎是由于受体存在可变磷酸化形式。在子宫切片中检查了磷酸化反应。在无激素的情况下,胞质受体被磷酸化。当添加激素时,受体的磷酸化明显增强,“核”受体的电泳迁移率降低。因此,这些实验表明“胞质”形式的受体是一种磷蛋白。在激素的作用下,受体在一些额外的位点进一步磷酸化。这种多磷蛋白是染色质结合的、推测具有活性的受体形式。在这方面,细胞内孕激素受体类似于各种激素和生长因子的膜受体,它们在结合配体后会被磷酸化。