转录因子AP-2α与TES蛋白相互作用分析

Analysis of interaction of AP-2α and TES

为研究转录因子AP-2α与TES蛋白的相互作用机制,分别构建AP-2α与GST的融合蛋白原核表达载体及TES与His的融合蛋白原核表达载体,在大肠杆菌E.coli BL 21中诱导表达,表达产物经纯化后用GST Pull-down及免疫印迹技术分析其相互作用关系.实验结果表明,AP-2α与TES之间存在相互作用是由于AP-2αC端能与TES C端的首个LIM结构域结合,酵母双杂交实验也证实了该结果;同时根据AP-2α只能与TES C端而不能与TES全长蛋白结合的实验事实提出了TES蛋白存在自身相互作用的可能,这种自身折叠封闭了位于TES C端首个LIM结构域上的AP-2α结合位点.

To study the interaction mechanism between transcription factor AP-2α and TES protein,prokaryotic vectors for expressing GST-AP-2α and His-TES were constructed respectively.Fusion proteins were induced in E.coli BL 21 and then were analyzed with GST pull-down assay.Results show that the interaction between TES and AP-2α is due to AP-2α C-terminus binding with the first LIM domain of TES C-terminus.These results were also confirmed in yeast two-hybrid system.Furthermore,a possible self-interaction of TES pro...