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目的探讨泛素连接酶Smad泛素化相关因子2(Smad ubiquitination-related factor 2,Smurf2)Nedd化修饰的机制。方法梯度过表达Nedd8检测Smuff2的蛋白水平变化,免疫共沉淀(IP)和Western印迹分析Smurf2的Nedd化修饰,GST融合蛋白沉降技术(GST-pulldown)验证相互作用,分析Smurf1 C426A基因敲入(knock in)小鼠体内Smurf2在淋巴、脾、肝和肺组织的蛋白表达水平。结果 Smurf2可发生Nedd化修饰,过表达Nedd8可导致Smurf2蛋白水平下调,Smurf1和Smurf2能相互作用,且Smurf1可使Smurf2的Nedd化修饰增强。在Smurf1 C426A基因敲入小鼠的组织细胞中Smurf2蛋白水平上调。结论 Smurf1能促进Smurf2的Nedd化修饰。
Objective To investigate the mechanism of Neddylation of Smad ubiquitination-related factor 2 (Smurf2). Methods Smurf1 C426A gene knock-in (knock-in) was detected by Western blotting analysis of Smed2, Nedd8, Nedd8, Nedd8 and GST-pulldown. in mice in vivo expression of Smurf2 protein in the lymph, spleen, liver and lung tissue. Results Smurf2 could be modified by Nedd. Overexpression of Nedd8 led to down-regulation of Smurf2 protein, Smurf1 and Smurf2 interaction, and Smurf1 enhanced Nurd2 modification. Smurf2 protein levels are up-regulated in the tissue cells of Smurf1 C426A knock-in mice. Conclusion Smurf1 can promote the Neddylation of Smurf2.