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为研究衰变加速因子 (DAF )介导Jurkat细胞信号传递的机制 ,本文应用免疫印迹及化学发光技术 (ECL)发现交联DAF单抗可使CD3+ 与CD3 Jurkat细胞总蛋白酪氨酸磷酸化水平增加 ,但磷酸化水平不同。免疫共沉淀实验检测到DAF与Src家族酪氨酸激酶 (SrcPTK )可发生共沉淀反应。去污剂不溶膜复合物 (DIG )抽提与鉴定检测到DIG内有SrcPTK与DAF的特异性聚集。说明DAF对Jurkat细胞的信号传递有辅助激活效应 ,此效应可能是通过其以与膜微区相关的所联系的SrcPTK实现的
In order to study the mechanism of decay accelerating factor (DAF) -mediated Jurkat cell signaling, we found that the cross-linked DAF McAb can increase the total protein tyrosine phosphorylation of CD3 + and CD3 Jurkat cells by Western blotting and chemiluminescence (ECL) , But different levels of phosphorylation. Co-precipitation reaction of DAF with Src family tyrosine kinase (SrcPTK) was detected by co-immunoprecipitation assay. Detergent insoluble membrane complex (DIG) extraction and identification detected specific aggregation of SrcPTK and DAF within the DIG. Indicating that DAF has an auxiliary activating effect on the signaling of Jurkat cells, possibly by the associated SrcPTK associated with the membrane microdomains