PTD-Hsp27 융합 단백질의 치료 효과 및 안정성 향상에 관한 연구

Abstract

Protein transduction domains (PTDs), also known as cell penetrating peptides (CPPs) are short peptide of amino acids which can translocate across cell membrane. Cargo, like genes, proteins and polymers, can be transferred efficiently into cells through several mechanism of PTD. Among all PTDs, transactivator of transcription (Tat) peptide derived from HIV-1 is widely used for transduction domain. Although mechanism of intracellular transduction of Tat is not well understood yet, it is considered that arginine of Tat peptide which has positive charge is a crucial factor for interaction with cell membrane. Therefore, nona-arginine (9R) was developed as effective peptide for transduction domain which can interact with negative charged phosphate at cell membrane. In this study, to confirm the efficient transduction efficacy for delivery of therapeutic protein (Heat shock protein 27; Hsp27) various combination of PTDs were constructed with Tat and 9R at N-terminal and C-terminal of Hsp27. PTD-Hsp27 was characterized by electrophoresis and visualized through confocal laser scanning microscopy (CLSM). To verify the enhanced effect of transduction by PTD conjugation, Cell viability was measured in hypoxic condition. Through this study, it is suggested that Tat peptide is optimum PTD for delivery of Hsp27.