基于生物信息学的抑癌基因NDRG2相互作用蛋白预测及验证

Bioinformatic Prediction and Experimental Validation of NDRG2 Interaction Partners

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. Although previous studies suggested that NDRG2 was implicated in tumor suppression and cellular differentiation, its intracellular signaling pathway and regulatory mechanism remained to be elucidated. In the present study, NDRG2 interacting proteins were predicted and analyzed by interologs （orthologous interactions from other species） bioinformatic approach. Binding of NDRG2 with candidate interaction partners was confirmed by Co-immunoprecipitation （Co-IP） and His pull-down. A number of candidate NDRG2- interacting partners were identified by bioinformatic approach. Based on previous research findings, Gnbl, Rgsl6 and Rgs5 were selected for further experimental validation. The results of Co-IP and His pull-down showed that Rgs5 interacted with NDRG2, while Gnbl and Rgsl6 can not. Collectively, our results indicated that bioinformatics analysis combined with laboratory validation is a kind of high efficient research strategies. Using this strategy, we confirmed the interaction between NDRG2 and Rgs5 proteins. Moreover, our data provide effective clues for subsequent function research.