9-Polylysine protein transduction domain: Enhanced penetration efficiency of superoxide dismutase into mammalian cells and skin

Jinseu Park, Jiyoon Ryu, Li Hua Jin, Jae Hoon Bahn, Ju A. Kim, Chang Sik Yoon, Dae Won Kim, Kyu Hyung Han, Won Sik Eum, Hycok Yil Kwon, Tae Cheon Kang, Moo Ho Won, Jung Hoon Kang, Sung Woo Cho, Soo Young Choi

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

Antioxidant enzymes, such as superoxide dismutase (SOD) and catalase (CAT), have been considered to have a beneficial effect against various diseases that are mediated by the reactive oxygen species (ROS). Although a variety of modified recombinant antioxidant enzymes have been generated to protect against oxidative stresses, the lack of their transduction ability into cells resulted in a limited ability to detoxify intracellular ROS. To render the SOD enzyme capable of detoxifying intracellular ROS when added extracellularly, cell-permeable recombinant SOD proteins were generated. A human Cu,Zn-superoxide dismutase (Cu,Zn-SOD) gene was fused with a gene fragment that encodes the 9 amino acids Tat protein transduction domain (RKKRRQRRR) of HIV-1 and lysine rich peptide (KKKKKKKKK) in a bacterial expression vector in order to produce a genetic in-frame Tat-SOD and 9Lys-SOD fusion protein, respectively. The expressed and purified Tat-SOD and 9Lys-SOD fusion proteins can transduce into human fibroblast cells, and they were enzymatically active and stable for 24 h. The cell viability of the fibroblast cells that were treated with paraquat, an intracellular Superoxide anion generator, was increased by the transduced Tat-SOD or 9Lys-SOD. The transduction efficacy of 9Lys-SOD was more efficient than that of Tat-SOD. We evaluated the ability of the SOD fusion proteins to transduce into animal skin. This analysis showed that Tat-SOD and 9Lys-SOD fusion proteins efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin (judged by the immunohistochemistry and specific enzyme activities). The enzymatic activity of the transduced 9Lys-SOD was higher than that of Tat-SOD, indicating that the penetration of 9Lys-SOD was more efficient when put into the skin. These results suggest Tat-SOD and 9LyS-SOD fusion proteins can be used as anti-aging cosmetics, or in protein therapy, for various disorders that are related to this antioxidant enzyme and ROS.

Original languageEnglish (US)
Pages (from-to)202-208
Number of pages7
JournalMolecules and Cells
Volume13
Issue number2
StatePublished - Apr 2002
Externally publishedYes

Keywords

  • 9Lysine-SOD
  • HIV-1 Tat
  • Penetration
  • Protein Transduction Domain
  • Skin

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of '9-Polylysine protein transduction domain: Enhanced penetration efficiency of superoxide dismutase into mammalian cells and skin'. Together they form a unique fingerprint.

Cite this