Dok1与表皮生长因子受体之间相互作用的酵母双杂交研究  

Interaction between epidermal growth factor receptor with dok1by yeast two hybrid method

作　　者：张勇[1] 刘秀杰[1] 张骐[1] 阎志勇[1] 矫力[1] 刘翾[1] 路长林[1] 何成[1] 

机构地区：[1]第二军医大学基础医学部神经生物学教研室,上海200433

出　　处：《第二军医大学学报》2004年第7期755-758,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金 ( 3 0 0 70 167;3 0 0 2 80 0 3 ) ;上海市科技发展基金 ( 0 1JC14 0 0 4) ;全军医药卫生科研"十五"规划课题( 0 1J0 11) .

摘　　要：目的 :研究 downstream of kinases(dok1 )与表皮生长因子受体 (EGFR)之间的相互作用 ,寻找 EGFR的可能底物或调控蛋白 ,以深入认识 EGFR下游信号转导机制。 方法 :将 EGFR胞内域与 L ex A蛋白融合作为 DNA结合蛋白 ,分别将dok1、dok1 PTB及 dok1 ΔPTB与 B4 2 AD蛋白融合作为激活域蛋白 ,共转化酵母菌后 ,通过 β-半乳糖苷酶活性检测、生长实验以及免疫共沉淀实验分析它们之间的相互作用。结果 :dok1及 dok1 PTB与 EGFR之间在酵母中存在结合 ,而 dok1Δ PTB不与 EGFR结合。 结论 :首次证实 EGFR与 dok1之间具有相互作用 ,dok 1的Objective:To study the interaction between epidermal growth factor receptor(EGFR)with downstream of kinases(dok1)and to search for the possible intracellular substrates or regulatory proteins of EGFR,so as to deeply under-stand the mechanisms of receptor downstream signal transduction.Methods:The EGFR IC (intracellular part of EGFR)was fused to LexA as a DNA-binding domain protein,while dok1,dok1PTB and dok1ΔPTB(dok1without PTB)were respective-ly fused to B42AD as an activation domain protein.Co-transformants were tested by β-galactosidase activity analysis,leucine medium growth analysis and co-immunoprecipitation assay.Results:Dok1and dok1PTB interacted with EGFR in yeast,while dok1ΔPTB did not.Conclusion:It is firstly confirmed that EGFR can interact with dok1,and the interaction may be mediated by PTB domain of dok1.

关 键 词：表皮生长因子受体 dok1 PTB 酵母双杂交 

分 类 号：R338[医药卫生—人体生理学]