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UFM化修饰在心肌肥厚小鼠中的变化及泛素折叠修饰因子1结合蛋白的筛选

UFMylation and Screening of Binding Proteins of Ubiquitin-fold Modifier 1 in Cardiac Hypertrophy
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摘要 目的 探讨UFM化修饰在心肌肥厚小鼠中的变化,并筛选潜在泛素折叠修饰因子1(UFM1)结合蛋白。方法 取8周龄C57BL/6J小鼠,背部皮下注射5 mg/(kg·d)的异丙肾上腺素(ISO)构建心肌肥厚模型。采用免疫荧光染色检测UFM1的表达水平及定位情况。采用蛋白免疫印迹法检测肥厚心肌组织中UFM化修饰变化及相关酶UFC1和UFL1的表达。通过免疫沉淀联合蛋白质谱筛选肥厚心肌组织内UFM1的结合蛋白。结果 在ISO诱导的心肌肥厚小鼠模型中,UFM1的表达水平显著增加。在肥厚心肌组织内,UFM化修饰水平升高,且伴有UFC1和UFL1表达水平的下调,提示心肌肥厚时UFM化修饰调控紊乱。采用蛋白质组学分析,共筛选到21种潜在的UFM1结合蛋白,其中FABP5、TNNT2、NCAM1、S100A8等可能介导了UFM化修饰与心肌肥厚之间的关联。结论 UFM化修饰在心肌肥厚小鼠中被激活,可能参与心肌肥厚发病的调控。 Objective To explore the status of UFMylation in cardiac hypertrophy and to screen potential binding proteins of ubiquitin-fold modifier 1(UFM1).Methods Eight-week-old C57BL/6J mice were injected back subcutaneously with 5 mg/(kg·d) of isoproterenol(ISO) to establish the cardiac hypertrophy model.The expression level and localization of UFM1 were detected by immunofluorescence staining.The changes of UFMylation and the expression of related enzymes,UFC1 and UFL1,in hypertrophic myocardial tissues were detected by Western blot.The binding proteins of UFM1 in hypertrophic myocardial tissues were screened by immunoprecipitation combined with protein mass spectrometry.Results The expression level of UFM1was significantly increased in the ISO-induced cardiac hypertrophy mouse model.In hypertrophic myocardium,the level of UFMylation was increased,whereas the levels of UFC1 and UFL1 were decreased,suggesting that the regulation of UFMylation was disordered in hypertrophic myocardium.Through proteomic analysis,21 potential UFM1 binding proteins were screened,among which FABP5,TNNT2,NCAM1,S100A8 may mediate the association between UFMylation and cardiac hypertrophy.Conclusions UFMylation is activated in cardiac hypertrophy and may be involved in the regulation of cardiac hypertrophy.
作者 冯景泓 赵晓涛 杨子瑶 孙媛媛 苏明 FENG Jinghong;ZHAO Xiaotao;YANG Ziyao;SUN Yuanyuan;SU Ming(Department of Laboratory,Peking University People’s Hospital,Beijing 100044,China)
出处 《中国分子心脏病学杂志》 CAS 2022年第6期5051-5056,共6页 Molecular Cardiology of China
基金 国家自然科学基金(81870196) 北京市自然科学基金(5192022)。
关键词 心肌肥厚 UFM化修饰 泛素折叠修饰因子1 Cardiac hypertrophy UFMylation Ubiquitin-fold modifier 1
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