Martin M, Böl G F, Eriksson A, Resch K, Brigelius-Flohé R
Medical School Hannover, FRG.
Eur J Immunol. 1994 Jul;24(7):1566-71. doi: 10.1002/eji.1830240717.
We have investigated the possibility of a protein kinase participating in the signal transduction mechanisms of the interleukin-1 (IL-1) type I receptor (IL-1RI). Our data show that a protein kinase was co-precipitated with the IL-1RI from the two murine T helper cell lines D10N and EL-4. The kinase activity was detected in an in vitro kinase assay performed with the immuno beads in the presence of exogenous substrates. IL-1 treatment of the cells resulted in a rapid activation of this protein kinase in a concentration-dependent manner. Both forms of IL-1, IL-1 alpha and IL-1 beta, induced this kinase activity, whereas the IL-1 receptor antagonist (IL-1ra) was inactive. In excess IL-1ra competitively antagonized IL-1 stimulation. In the in vitro kinase assay the exogenous substrates myelin basic protein and histone H1 were phosphorylated, whereas casein or heat-shock protein HSP27 were not accepted, reflecting a certain selectivity of this protein kinase. The IL-1RI co-precipitable protein kinase showed a serine/threonine specificity and was inhibited by staurosporine, but not by inhibitors specific for protein tyrosine kinase or protein kinase C. These results show that a serine/threonine protein kinase directly interacts with the IL-1RI at the plasma membrane level of T helper cells forming a novel type of IL-1 inducible signaling complex. This protein kinase may resemble the link coupling the plasma membrane IL-1 receptor to cytosolic downstream elements in the IL-1 signaling pathway.
我们研究了蛋白激酶参与白细胞介素-1(IL-1)I型受体(IL-1RI)信号转导机制的可能性。我们的数据表明,在两种小鼠T辅助细胞系D10N和EL-4中,一种蛋白激酶与IL-1RI共沉淀。在用免疫珠在存在外源底物的情况下进行的体外激酶测定中检测到了激酶活性。用IL-1处理细胞导致该蛋白激酶以浓度依赖的方式快速激活。两种形式的IL-1,即IL-1α和IL-1β,均可诱导这种激酶活性,而IL-1受体拮抗剂(IL-1ra)则无活性。过量的IL-1ra竞争性拮抗IL-1刺激。在体外激酶测定中,外源底物髓鞘碱性蛋白和组蛋白H1被磷酸化,而酪蛋白或热休克蛋白HSP27未被磷酸化,这反映了该蛋白激酶具有一定的选择性。与IL-1RI共沉淀的蛋白激酶表现出丝氨酸/苏氨酸特异性,并且被星形孢菌素抑制,但不受蛋白酪氨酸激酶或蛋白激酶C特异性抑制剂的抑制。这些结果表明,一种丝氨酸/苏氨酸蛋白激酶在T辅助细胞的质膜水平与IL-1RI直接相互作用,形成一种新型的IL-1诱导信号复合物。这种蛋白激酶可能类似于将质膜IL-1受体与IL-1信号通路中的胞质下游元件偶联的连接物。