【摘要】 目的寻找人类14-3-3ζ蛋白的相互作用蛋白,为进一步揭示14-3-3ζ的作用机理提供线索。方法以14-3-3ζ为"诱饵",利用酵母双杂交系统筛选人脑cDNA文库得到"猎物"蛋白,通过GST pull-down技术和哺乳动物细胞内免疫共沉淀技术验证14-3-3ζ和"猎物"蛋白的特异性结合。结果首次利用酵母双杂交cDNA文库筛选技术发现了14-3-3ζ能够与GTP环化水解酶Ⅰ(GTP cyclohydrolase1,GCH1)相互作用,并通过了体内和体外的蛋白质结合实验证实了这两个蛋白质之间的特异性相互作用。结论发现并验证了14-3-3ζ与GCH1之间的蛋白质相互作用,为进一步深入研究这两种蛋白质的功能及所引起的相关疾病的发病机理提供了新的线索。更多还原

【Abstract】 Objective To search for proteins interacting with human 14-3-3ζ in order to provide further clues to the mechanism of action of 14-3-3ζ.Methods We used 14-3-3ζ as "bait" to screen a human brain cDNA library,obtained "prey" proteins,then confirmed the interaction by GST pull-down assay in vitro and co-immunoprecipitation assay in vivo.Results We found that 14-3-3ζ interacted with GCH1(GTP cyclohydrolase 1)by using yeast two-hybrid method,and the specific interaction between these two proteins was confirmed by protein binding experiments both in vitro and in vivo.Conclusion Our study verifies the protein interaction between 14-3-3ζ and GCH1,and provides a new clue to the further research of functions of these two proteins and the mechanism of the related diseases.更多还原