标题: 建构Securin蛋白质交互作用网络
Construction of Securin Interaction Network
作者: 杨瑶贞
张家靖
Chang, Chia-Ching
分子医学与生物工程研究所
关键字: 网络;远西方墨点法;securin;network;Far Western
公开日期: 2008
摘要: Securin是一个多功能蛋白质,且在许多癌症中有高量的表现,但是对于securin如何调控癌症细胞的机制目前尚未完全解开,所以希望能藉由建构与securin蛋白质作用的网络,更进一步的了解securin的作用机制。我们使用免疫沉淀(IP)、酵母菌双杂交系统(yeast two hybrid)及远西方墨点法(Far Western)等方法筛选出与securin结合的蛋白质,经由质谱仪(MALDI-TOF Mass)分析其序列后,确定这些蛋白质为PCAF、T-box3、Histone H4、KRT1、BCl2A1、Dopamine receptor D4、及RNA cyclase,藉由这些与securin作用的蛋白质,我们可以知道,securin除了已知功能外,还可能与蛋白质乙醯化的调节、Histone H4、Cytoskeleton及抗细胞凋亡的BCl2A1有关。另外我们使用两种方式测量蛋白质之间的交互作用:远西方墨点(Far Western)法及恒温滴定微卡洛里计(ITC),实验中以p53与securin为代表,分别用两种方式测得其作用力大小,此测量系统能应用于测量不同环境下蛋白质之间的作用力,以及不同修饰状态下的蛋白质作用力大小,作为探讨蛋白质之间分子作用机制的分析方法。
Human securin is a multifunctional protein and highly expressed in many types of tumor cell. The function of securin is regulating separase that control cell division and cell cycle, and DNA repair mechinary. However, the molec1ule mechanism of securin in cell cycle regulation remains unclear. To understand the role of securin in cell cycle, a protein-protein interaction network of securin was constructed try using immunoprecipitation, yeast two hybrid and Far Western methods. We found several proteins may interact with securin such as PCAF, T-box3, Histone H4, KRT1, BCl2A1, Dopamine receptor D4 and RNA cyclase. From the results of the experiments, securin may participate in protein acetylation, mofification of Histone H4 and apoptosis. Meanwhile we measured the protein-protein interaction by using Far Western and ITC. The two systems offered us a useful tool to analyze protein-protein interaction in different environments and different situations of protein modification.
URI: http://140.113.39.130/cdrfb3/record/nctu/#GT079629509
http://hdl.handle.net/11536/42741
显示于类别:Thesis


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