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[Cell-penetrating peptide-based functional study of p38 MAPK].
Li-ping Yang1, Zhi-feng Liu, Yong-ming Li
1Department of Pathophysiology and Guangdong Provincial Key Laboratory of Functional Proteomics, Southern Medical University, Guangzhou 510515, China.
Summary
A novel TAT protein-based delivery system successfully introduced p38 mitogen-activated protein kinase (MAPK) into eukaryotic cells. This system effectively modulated endogenous p38 activity, with implications for signal transduction research.
Area of Science:
- Cellular biology
- Molecular biology
- Signal transduction
Context:
- The p38 MAPK pathway plays a crucial role in cellular responses to stress.
- Efficient delivery of functional proteins into eukaryotic cells remains a challenge.
- The TAT protein is known for its cell-penetrating capabilities.
Purpose:
- To develop a protein delivery system for p38 MAPK using the TAT protein.
- To investigate the intracellular function and biological activity of delivered p38 MAPK.
- To assess the impact of TAT-mediated p38 MAPK delivery on endogenous signaling pathways.
Summary:
- Recombinant vectors encoding His-TAT-p38 and a phosphorylation-deficient mutant (His-TAT-p38(AF)) were constructed and expressed in E. coli.
- Purified His-TAT-p38 and His-TAT-p38(AF) proteins were efficiently delivered into ECV304 cells in a time- and concentration-dependent manner.
- His-TAT-p38 enhanced endogenous p38 activity, while His-TAT-p38(AF) inhibited ATF2 phosphorylation, blocking the p38 signaling pathway under osmotic stress.
Impact:
- Successful construction of a TAT protein-based p38 MAPK delivery system.
- Demonstrated efficient, dose-dependent cellular uptake of TAT-fused proteins.
- Established the biological activity of delivered p38 MAPK and its mutant in modulating cellular signaling pathways.