非受体酪氨酸激酶c-Abl与Siva-1相互作用及在细胞凋亡中的功能研究  

Interaction between non-receptor tyrosine kinase c-Abl and Siva-1 in cell apoptosis

作　　者：李平[1] 李楚芳[1] 刘萱[1] 靳彦文[1] 黄维[1] 易艳平[1] 马清钧[1] Donald W Kufe 曹诚[1] 

机构地区：[1]军事医学科学院生物工程研究所 [2]Dana Farber Cancer Institute,Harvard Medical School,MA02115,USA

出　　处：《军事医学科学院院刊》2003年第4期251-254,共4页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的 :研究非受体酪氨酸激酶c_Abl与Siva_1蛋白的相互作用及其对细胞凋亡的影响。方法 :应用免疫共沉淀研究蛋白质相互作用 ;抗酪氨酸抗体研究蛋白体内磷酸化 ;体外激酶分析研究体外磷酸化 ;流式细胞仪测定SubG1 细胞百分比以测定细胞调亡。结果 :非受体酪氨酸激酶c_Abl在细胞内及体外均与Siva_1形成复合物 ,通过体外翻译的c_Abl结合实验表明其为直接的相互作用 ,其相互作用主要通过c_Abl的SH3结构域与Siva_1直接结合 ,且Siva_1可以被c_Abl磷酸化。通过c_Abl显性负突变结果表明 ,酪氨酸磷酸化对Siva_1诱导细胞凋亡至关重要。结论 :c_Abl使Siva_1磷酸化可促进Siva_1的细胞凋亡功能。Objective:To demonstrate the interaction between non_receptor tyrosine kinase c_Abl and apoptosis inducing protein Siva_1. Methods:Interaction was demonstrated by immunoprecipitation, in vivo tyrosine phosphorylation was analyzed by immunoblotting with anti_phosphorylated tyrosine antibody, in vitro phosphorylation was analyzed by in vitro kinase assay and apoptosis was evaluated by the percentage of sub G 1 cells by FACScan.Results:Interaction of c_Abl with Siva_1 was demonstrated by immuno_coprecipitation and direct binding analysis. c_Abl bound to Siva_1 by its SH3 domain. Siva_1 could be phosphorylated by c_Abl in vitro or in vivo . Siva_1 cannot induce apoptosis in c_Abl(K_R) dominant negative cell line. Conclusion:Phosphorylation of Siva_1 by c_Abl plays an important role in Siva_1 induced apoptosis. [

关 键 词：C-ABL Siva-1 细胞凋亡 磷酸化 机制 

分 类 号：R73-3[医药卫生—肿瘤]