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亚溶解型C5b-9刺激GMC诱导KLF4与PCAF结合及结合部位的鉴定
作者:虞天一1  何风霞2 3  赵晨卉4  余明皓3  赵聃3  刘玉3  宫雅娟3  夏露3  邱文3  张婧3  王迎伟3 
单位:1. 东南大学附属中大医院 妇产科, 江苏 南京 210009;
2. 南京医科大学附属第二医院 病理科, 江苏 南京 210003;
3. 南京医科大学 免疫学系, 江苏 南京 211166;
4. 南京医科大学第一附属医院 肿瘤科, 江苏 南京 210029
关键词:亚溶解型C5b-9 大鼠肾小球系膜细胞 Kruppel样因子4 P300/CBP相关因子 结合部位 
分类号:R392
出版年·卷·期(页码):2018·37·第六期(969-973)
摘要:

目的:鉴定亚溶解型C5b-9刺激大鼠肾小球系膜细胞(GMC)诱导Kruppel样因子4(KLF4)与P300/CBP相关因子(PCAF)相互结合以及结合部位。方法:以亚溶解型C5b-9刺激GMC,应用免疫沉淀(IP)和免疫印迹(IB)法检测KLF4与PCAF相互结合情况。另构建KLF4、PCAF过表达和KLF4、PCAF不同结构域的质粒,将质粒转染GMC,应用IP和IB鉴定KLF4与PCAF的结合部位。结果与结论:亚溶解型C5b-9刺激GMC后能够诱导KLF4与PCAF相互结合,KLF4结合于PCAF转录结合结构域,而PCAF则结合在KLF4的转录激活结构域。

Objective:To identify kruppel-like factor 4(KLF4) binding to P300/CBP-associated factor (PCAF) and the binding site induced by sublytic C5b-9-stimulated rat glomerular mesangial cells (GMC). Methods:GMC were stimulated by sublytic C5b-9. Then KLF4 binding to PCAF were examined by immunoprecipitation (IP) and immunoblot (IB). Meantime, the plasmids of pIRES2-KLF4 and pIRES2-PCAF as well as structure-domain of KLF4 and PCAF were constructed. Then GMC were co-transfected by these plasmids to determine the binding sites between KLF4 and PCAF by IP and IB. Results and Conclusion:Sublytic C5b-9 can induce the combination of KLF4 and PCAF through stimulation GMC. KLF4 binding to PCAF located in the activation domain of KLF4, while the site of PCAF binding to KLF4 located in the transcriptional binding domain of PCAF.

参考文献:

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