摘要
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 Smad ubiquitination-related factor 2（Smurf2） neddylation.Methods The Smurf2 protein level was tested by overexpression of Nedd8,while the method of immunoprecipitation（IP）and Western blotting were used to analyz Smurf2-Nedd8 modification.The GST-pulldown experiment was conducted to prove protein interactions.The protein was obtained by grinding mouse tissue and Western blotting was used to test the protein expression level.Results Over expression of Nedd8 could lead to the down regulation of the SmurG＇s protein level.Smurfl and Smurf2 could interact in the GST-pulldown experiment.Smurf1 could enhance Smurf2-Nedd8 modification.The Smurf2＇s protein level was up-regulated in mouse tissue of Smurf1 C426 A.Conclusion There is some relationship and also difference between Smurf1 and Smurf2.Smurf1 can enhance Smurf2-Nedd8 modification.
作者
刘超
李海文
韦荣飞
谢萍
张令强
LIU Chao LI Hai-wen WEI Rong-fei XIE Ping ZHANG Ling-qiang(Graduate School, Anhui Medical University, Hefei 230032, China Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 100850, China Capital Medical University, Beijing 100730, China)
出处
《军事医学》
CAS
CSCD
北大核心
2017年第2期86-89,共4页
Military Medical Sciences
基金
北京市科技新星计划资助项目(Z16111000490000)
国家自然科学基金资助项目(31470035)
