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本文对增殖期的淋巴细胞胰岛素依赖性酪氨酸蛋白激酶活性及内源性废物进行了分析研究。在纯化的健康人淋巴细胞中加入适量的植物血凝素(PHA),经过72h培养即成为转化淋巴细胞(增殖期淋巴细胞)。应用~(32)P参入实验,证实转化淋巴细胞胰岛素受体具有胰岛素依赖性的酪氨酸蛋白激酶活性,与未转化的对照组相比其活性增加约9倍。Scatchard分析表明转化后淋巴细胞膜表面胰岛素受体数增加3.5倍。应用抗酪氨酸磷酸酯抗体,对胰岛素作用前后的转化与未转化淋巴细胞内,酪氨酸残基磷酸化的蛋白进行了鉴定,结果表明:除了95kD受体β亚基自身磷酸化外,45kD蛋白质也明显磷酸化,我们命名它为PP45。我们认为PP45可能是淋巴细胞中胰岛素受体酪氨酸蛋白激酶的主要内源性废物,它的磷酸化是胰岛素信息传递过程级联反应的初始步骤。
In this paper, the proliferation of lymphocyte insulin-dependent tyrosine kinase activity and endogenous waste were analyzed. Purified healthy human lymphocytes by adding appropriate amount of phytohemagglutinin (PHA), after 72h culture to become transformed lymphocytes (proliferating lymphocytes). The application of ~ (32) P into the experiment, confirmed that transformed lymphocyte insulin receptor insulin-dependent tyrosine kinase activity, compared with the untransformed control group increased by about 9-fold. Scatchard analysis showed a 3.5-fold increase in the number of insulin receptors on the surface of transformed lymphocyte membranes. Application of anti-tyrosine phosphates antibody, before and after the conversion of insulin and non-transformed lymphocytes, tyrosine phosphorylation of protein were identified, the results showed that: in addition to 95kD receptor β subunit autophosphorylation, The 45kD protein is also clearly phosphorylated, and we name it PP45. We believe that PP45 may be the major endogenous waste of insulin receptor tyrosine kinase in lymphocytes and its phosphorylation is an initial step in the cascade of insulin signaling.