The J-domain of heat shock protein 40 can enhance the transduction efficiency of arginine-rich cell-penetrating peptides

热休克蛋白40的J结构域可以增强富含精氨酸的细胞穿透肽的转导效率

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作者:Tzu-Yin Lin, Yu-Hsiu Su, Kun-Hsiung Lee, Chin-Kai Chuang

Abstract

Sense and antisense oligonucleotide pairs encoding cell-penetrating peptides PTD (Tat47-57), DPV3A, E162, pVEC, R11, and TP13 were used to construct two sets of pET22b-CPP-DsRed and pET22b-CPP-J-DsRed vectors for CPP-DsRed and CPP-J-DsRed recombinant proteins expression. PTD-DsRed, DPV3A-DsRed, PTD-J-DsRed, and DPV3A-J-DsRed recombinant proteins were expressed in a soluble form. PTD-J-DsRed and DPV3A-J-DsRed recombinant proteins were able to escape from E. coli host cells into the culture medium. The membrane-penetrating activity of PTD-J-DsRed and DPV3A-J-DsRed recombinant proteins to mammalian cells was more effective than that of PTD-DsRed and DPV3A-DsRed. The route of the cellular membrane translocation of these recombinant proteins is suggested via macropinocytosis followed by an endosomal escape pathway.

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