Interaction of the insulin receptor kinase with serine/threonine kinases in vitro.
Interaction of the insulin receptor kinase with serine/threonine kinases in vitro.
复制标题
体外胰岛素受体激酶与丝氨酸/苏氨酸激酶的相互作用。
DOI：
10.1002/jcb.240280209
复制
发表时间：
1985
影响因子：
4
通讯作者：
Roach,PJ
中科院分区：
文献类型：
--
作者：
Haring,HU;White,MF;Kahn,CR;Ahmad,Z;DePaoli-Roach,AA;Roach,PJ
在破碎的细胞制备物中，胰岛素引起其受体β亚基（Mr = 95，000）的快速磷酸化。这发生在酪氨酸残基上，并且是由于受体本身所含的蛋白激酶的激活。在完整细胞中，胰岛素还刺激丝氨酸和苏氨酸残基上的受体和其他细胞蛋白的磷酸化。在试图找到一种蛋白质，可能连接受体酪氨酸激酶这些丝氨酸/苏氨酸磷酸化反应，我们已经研究了部分纯化的制剂的胰岛素受体与纯化制剂的丝氨酸/苏氨酸激酶已知磷酸化糖原合酶的相互作用。当酪蛋白激酶I和II、磷酸化酶激酶或糖原合成酶激酶3与胰岛素受体体外孵育时，未观察到胰岛素依赖性磷酸化。这些激酶也不能使受体磷酸化。相比之下，胰岛素受体激酶催化钙调蛋白依赖性激酶的磷酸化，体外添加胰岛素导致磷酸化增加40%。在钙调蛋白依赖性激酶和胰岛素受体激酶存在的情况下，胰岛素也刺激钙调蛋白的磷酸化。磷酸化氨基酸分析显示钙调素和钙调素非依赖性蛋白激酶磷酸化酪氨酸含量增加。这些数据表明，胰岛素受体激酶可能直接和特异性地与钙调蛋白依赖性激酶和钙调蛋白相互作用。需要进一步的研究来确定这些磷酸化是否改变了这些调节蛋白的作用。
Insulin causes rapid phosphorylation of the β subunit (Mr= 95,000) of its receptor in broken cell preparations. This occurs on tyrosine residues and is due to activation of a protein kinase which is contained in the receptor itself. In the intact cell, insulin also stimulates the phosphorylation of the receptor and other cellular proteins on serine and threonine residues. In an attempt to find a protein that might link the receptor tyrosine kinase to these serine/threonine phosphorylation reactions, we have studied the interaction of a partially purified preparation of insulin receptor with purified preparations of serine/threoine kinases known to phosphorylate glycogen synthase. No insulin‐dependent phosphorylation was ob served when casein kinases I and II, phosphorylase kinase, or glycogen synthase kinase 3 was incubated in vitro with the insulin receptor. These kinases also failed to phosphorylate the receptor. By contrast, the insulin receptor kinase catalyzed the phosphorylation of the calmodulin‐dependent kinase and addition of insulin in vitro resulted in a 40% increase in this phosphorylation. In the presence of calmodulin‐dependent kinase and the insulin receptor kinase, insulin also stimulated the phosphorylation of calmodulin. Phosphoamino acid analysis showed an increase of phosphotyrosine content in both calmodulin and calmodulindependent protein kinase. These data suggest that the insulin receptor kinase may interact directly and specifically with the calmodulin‐dependent kinase and calmodulin. Further studies will be required to determine if these phosphorylations modify the action of these regulatory proteins.