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NDRG2(N-Myc downstream regulator gene 2)是NDRG家族成员之一.以往研究表明,该家族与细胞的增殖和分化有关.而该分子参与的细胞信号通路及调节机制尚未阐明.本研究利用保守蛋白间相互作用(interologs)的生物信息学方法预测NDRG2相互作用分子,并通过免疫共沉淀(Co-IP)及His pull-down蛋白体外结合实验方法对预测结果进行验证.生物信息学软件预测和分析结果表明,细胞中存在多个可能与NDRG2发生相互作用分子.结合文献报道,从中选取了3个候选分子Gnb1、Rgs16及Rgs5进行分子生物学实验验证.Co-IP及His pull-down实验结果表明,3个候选分子中,Rgs5蛋白能够和NDRG2蛋白相互作用,而其它2个候选分子与NDRG2的相互作用未获得实验室方法的验证.研究结果表明,生物信息学分析与实验室验证相结合是一种高效省时的蛋白质相互作用研究策略.通过这种策略证实NDRG2可以与Rgs5蛋白相互作用,为后续NDRG2功能的研究提供了有效的线索.
NDRG2 (N-Myc downstream regulator gene 2) is a member of the NDRG family.Researches have shown that this family is related to the proliferation and differentiation of cells, and the molecular signaling pathway and regulatory mechanisms involved in this molecule have not been elucidated.In this study, The bioinformatics methods of interologs predicted the NDRG2 interacting molecules and validated the predictive results by co-immunoprecipitation (Co-IP) and His pull-down protein in vitro binding assays. Bioinformatics Software Prediction and Analysis The results showed that there were many molecules that may interact with NDRG2 in the cell.We selected three candidate molecules Gnb1, Rgs16 and Rgs5 for molecular biology experiments in the literature.Co-IP and His pull-down experiments showed that , Three candidate molecules, Rgs5 protein and NDRG2 protein interaction, and the other two candidate molecules and NDRG2 interaction has not been verified by laboratory methods.The results show that the combination of bioinformatics analysis and laboratory verification is A highly efficient and time-saving protein-protein interaction strategy, through which NDRG2 interacts with Rgs5 protein The study of NDRG2 function provides effective clues.