Protein kinase activity associated with the purified rat hepatic glucocorticoid receptor.
Protein kinase activity associated with the purified rat hepatic glucocorticoid receptor.
复制标题
与纯化的大鼠肝糖皮质激素受体相关的蛋白激酶活性。
DOI：
10.1073/pnas.82.12.4003
复制
发表时间：
1985
影响因子：
11.1
通讯作者：
Litwack,G
中科院分区：
文献类型：
--
作者：
Miller-Diener,A;Schmidt,TJ;Litwack,G
纯化5000倍的大鼠肝糖皮质激素受体的Mr 94，000类固醇结合组分在体外通过[γ-32 P]ATP进行钙刺激磷酸化。外源性组蛋白可以被这种无钙制剂磷酸化。钙调素没有刺激糖皮质激素受体的磷酸化，超过了单独使用钙的情况。特异性钙调素抑制剂calimidazolium无作用，而非特异性钙调素抑制剂三氟拉嗪和氯丙嗪则能阻断受体的钙依赖性磷酸化。EGTA阻断钙的作用;镁不能代替钙。环核苷酸（cAMP或cGMP）在无钙的情况下不刺激受体的磷酸化。糖皮质激素受体的磷酸化是类固醇依赖性的。曲安奈德引起激活和磷酸化的受体存在的钙，而孕酮拮抗剂，皮质醇，β-拉帕酮没有。阻断热活化步骤的Na 2SO 4抑制受体的磷酸化。糖皮质激素受体的活化形式是磷酸化发生所必需的。ATP类似物8-叠氮基-ATP或氟磺酰基苯甲酰基腺苷抑制Mr 94，000组分的磷酸化，这意味着存在受体固有的ATP结合位点。
The Mr 94,000 steroid binding component of rat hepatic glucocorticoid receptor purified 5000-fold under-goes calcium-stimulated phosphorylation in vitro by [gamma-32P]ATP. Exogenous histones can be phosphorylated by this preparation without calcium. Calmodulin did not stimulate phosphorylation of the glucocorticoid receptor beyond that obtained with calcium alone. Although the specific calmodulin inhibitor calmidazolium had no effect, trifluoperazine and chlorpromazine, nonspecific calmodulin inhibitors, abolished the calcium-dependent phosphorylation of receptor. EGTA blocks the effect of calcium; magnesium cannot substitute for calcium. Cyclic nucleotides (cAMP or cGMP) do not stimulate phosphorylation of the receptor in the absence of calcium. Phosphorylation of the glucocorticoid receptor is steroid dependent. Triamcinolone acetonide elicited activation and phosphorylation of receptor in the presence of calcium, whereas the antagonists progesterone, cortexolone, and beta-lapachone did not. Sodium molybdate, which blocks the thermal activation step, inhibits phosphorylation of the receptor. The activated form of the glucocorticoid receptor is required for phosphorylation to occur. The ATP analogues 8-azido-ATP or fluorosulfonylbenzoyl adenosine, inhibit phosphorylation of the Mr 94,000 component, implying the presence of an ATP binding site inherent to the receptor.