PLC-β3与syntrophin蛋白的相互作用  

The interaction between PLC-β3 and syntrophin

作　　者：黄艳[1] 熊英[1] 刘华[1] 孙超渊[1] 李洋[1] 贺俊崎[1] 

机构地区：[1]首都医科大学基础医学院生物化学与分子生物学系,北京100069

出　　处：《基础医学与临床》2010年第7期708-713,共6页Basic and Clinical Medicine

基　　金：国家自然基金(30772573;30973405);北京市自然基金(7082009);北京市教育委员会科技发展计划重点项目(KZ201010025026);北京市"创新人才建设计划"项目

摘　　要：目的探索并鉴定PLC-β3与PDZ蛋白syntrophin的相互作用,为进一步研究PLC-β3在神经系统信号传导通路中的作用提供线索。方法制备GST-PLC-β3-CT融合蛋白,利用GST pull down的方法,检测PLC-β3-CT与β2-syntrophin和γ2-syntrophin的PDZ结构域的相互作用。结果构建重组质粒pGEX-PLC-β3-CT,在大肠杆菌BL21中成功表达融合蛋白后,利用GST pull down的方法证实了PLC-β3-CT可以与β2-syntrophin而不是γ2-syntrophin的PDZ结构域相互作用。结论 PLC-β3与β2-syntrophin相互作用,为研究完整的PLC-β3与β2-syntrophin的相互作用以及这种相互作用在神经系统信号网络中的功能提供了线索。Objective To identify the interaction between PLC-β3 and the PDZ domain-containing protein syntrophin,and to find the underlying roles of PLC-β3 in signaling pathway of the nervous system.Methods The carboxyl-terminal of PLC-β3（PLC-β3-CT） was fused to GST protein by gene cloning.The resultant protein was used for examining interactions of PLC-β3-CT with the PDZ-domain of β2-syntrophin and γ2-syntrophin respectively by GST pull-down assay.Results PLC-β3-CT cDNA was cloned into the expression vector pGEX-4T-1 successfully,and the GST fusion protein was overexpressed and purified in an E.coli system.GST pull-down experiments revealed that the PDZ domain of β2-syntrophin,but not γ2-syntrophin was robustly pulled down by the carboxyl-terminal of PLC-β3.Conclusion These results provide a strong evidence for further studying the intracellular association of the PLC-β3 and β2-syntrophin and for exploring their potential physiological significances in signaling pathway of the nervous system.

关 键 词：PLC-β3 SYNTROPHIN 蛋白相互作用 

分 类 号：Q26[生物学—细胞生物学]