首页|期刊导航|老年心脏病学杂志（英文版）|Functional study of p38 mitogen-activated protein kinase based on cell-penetrating peptide delivery system
老年心脏病学杂志（英文版）2009，Vol.6Issue(2)：108-114,7.
Functional study of p38 mitogen-activated protein kinase based on cell-penetrating peptide delivery system
Functional study of p38 mitogen-activated protein kinase based on cell-penetrating peptide delivery system
摘要
Abstract
Objective p38 Mitogen-activated protein kinase (MAPK) is a crossing center of various pathways. In this study, protein transduction system based on human immunodeficiency virus (HIV)-1 transactivator of transcription (TAT), which is an efficient delivery peptide of the foreign proteins into cells, was employed to study p38 MAPK functions in eukaryotic cells. Methods p38 And its dominant negative form, p38AF, were constructed into pET-His-TAT vector correctly to verify that the recombinant plasmids were well-founded through restriction enzyme digestion and DNA sequencing. The two proteins, His-TAT-p38 and His-TAT-p38AF, were expressed and purified in Escherichia coli by SDS-PAGE. Then they were incubated with ECV304 cells respectively and readily transduced into cells in a time-dependent and dose-dependent manner. The cells were stimulated by sorbitol. Activating transcription factor (ATF) 2 phosphorylation level was checked using Western blot to assess the activity of endogenous p38. Results Compared with controls, it was found that His-TAT-p38 increased the level ofATF2 phosphorylation in sorbitol-stimulated ECV304 cells, while His-TAT-p38AF inhibited it, indicating p38 MAPK protein delivery system based on TAT was constructed successfully. TAT-p38 and its dominant negative form possessed high biological activity after transduction into ECV304 cells by TAT protein delivery system. The results showed that p38AF fused with TAT could inhibit the transduction of endogenous p38 signal pathway in part, and other pathway might regulate p38 phosphorylation. Conclusions Our study provides a novel pathway to inhibit p38 signal pathway and establish a new method to study p38 function.关键词
Human immunodeficiency virus-1/transactivator of transcription/p38 mitogen-activated protein kinase/protein transduction/sorbitolKey words
Human immunodeficiency virus-1/transactivator of transcription/p38 mitogen-activated protein kinase/protein transduction/sorbitol分类
医药卫生引用本文复制引用
Liping Yang,Yongming Yao,Zhiyong Sheng,Xiaomei Zhu,Yong Jiang..Functional study of p38 mitogen-activated protein kinase based on cell-penetrating peptide delivery system[J].老年心脏病学杂志（英文版）,2009,6(2):108-114,7.基金项目
This work was supported,in part,by grants from the National Basic Research Program of China (2005CB522602),the National Natural Science Foundation (30672178,30872683,30800437),and the National Natural Science Outstanding Youth Foundation fo China (30125020). （2005CB522602）
