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음원식,In Soon Choung,A Yeon Kim,Yong Je Lee,Jung Hoon Kang,Jinseu Park,Kil Soo Lee,Hyeok Yil Kwon,최수영 한국분자세포생물학회 2002 Molecules and cells Vol.13 No.2
We previously reported that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. To enhance the therapeutic potential of Tat-SOD for the treatment of various disorders that are related to this antioxidant enzyme, the transduction efficacy of Tat-SOD should be heightened. Therefore, we investigated whether copper ion recovery of the fusion protein could enhance the transduction potential of Tat- SOD in cultured HeLa cells. The results showed that the transduction potential of Tat-SOD was markedly enhanced by copper ions, and moderately increased by zinc ions. Compared with Tat-SOD, the Tat-SOD that recovered the copper ion (CR-Tat-SOD) achieved a significant increase in intracellular concentration and enzymatic activity. Therefore, CR-Tat-SOD was transduced into HeLa cells in a rapid saturation manner, but Tat-SOD was shown in a time-dependent manner. With the higher transduction efficacy of CR-Tat-SOD than that of Tat-SOD, the transduced CR-Tat-SOD significantly increased the viability of HeLa cells that were pretreated with paraquat, an intracellular superoxide anion generator. Although the mechanism of the enhanced transduction of Tat-SOD by copper ions is still unanswered, these results indicate that copper ions facilitate the transduction of SOD. These then significantly increase the biological effectiveness of this antioxidant enzyme.
박진서,최수영,음원식,장상호,김대원,최희순,Soo Hyun Choi,So Young Kim,Jae Jin An,Sun Hwa Lee,한규형,Jung Hoon Kang,강태천,원무호,Yong Joon Cho,Jin Hi Choi,Tae Yoon Kim 한국분자세포생물학회 2005 Molecules and cells Vol.19 No.2
The human immunodeficiency virus type 1 (HIV-1) Tat protein transduction domain (PTD) is responsible for highly efficient protein transduction across plasma membranes. In a previous study, we showed that Tat- Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. In this study, we fused the human SOD gene with a Tat PTD transduction vector at its N- and/or C-terminus. The fusion proteins (Tat-SOD, SOD-Tat, Tat-SOD-Tat) were purified from Escherichia coli and their ability to enter cells in vitro and in vivo compared by Western blotting and immunohistochemistry. The transduction efficiencies and biological activities of the SOD fusion protein with the Tat PTD at either terminus were equivalent and lower than the fusion protein with the Tat PTD at both termini. The availability of a more efficient SOD fusion protein provides a powerful vehicle for therapy in human diseases related to this anti-oxidant enzyme and to reactive oxygen species.
Transduced Tat-SOD Fusion Protein Protects Against Ischemic Brain Injury
김대원,최수영,음원식,장상호,김소영,최희순,최수현,안재진,이선화,이길수,한규형,강태천,원무호,강정훈,권오신,조성우,김태윤,박진서 한국분자세포생물학회 2005 Molecules and cells Vol.19 No.1
Reactive oxygen species (ROS) are implicated in reper-fusion injury after transient focal cerebral ischemia. The antioxidant enzyme, Cu,Zn-superoxide dismutase (SOD), is one of the major means by which cells coun-teract the deleterious effects of ROS after ischemia. Re-cently, we reported that when Tat-SOD fusion protein is transduced into pancreatic cells it protects the cells from destruction by relieving oxidative stress in ROS-implicated diabetes (Eum et al., 2004). In the present study, we investigated the protective effects of Tat-SOD fusion protein against neuronal cell death and ischemic insults. When Tat-SOD was added to the culture me-dium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Immunohistochemical analysis revealed that Tat-SOD injected intraperitoneally (i.p.) into mice has access to various tissues including brain neurons. When i.p. in-jected into gerbils, Tat-SOD prevented neuronal cell death in the hippocampus in response to transient fore-brain ischemia. These results suggest that Tat-SOD pro-vides a strategy for therapeutic delivery in various hu-man diseases, including stroke, related to this anti-oxidant enzyme or to ROS.
Penetration of a Tat-Superoxide Dismutase Fusion Protein into Skin
최수영 ( Soo Young Choi ),( Li Hua Jin ),( Jae Hoon Bahn ),( A Yeon Kim ),( Hyo Seop Kim ),( Joo A Kim ),( Yong Je Lee ),( Sang Ho Jang ),( Ji Yoon Ryu ),( Jin Seu Park ),( Won Sik Eum ),( Hyeok Yil Kwon 한국피부장벽학회 2001 한국피부장벽학회지 Vol.3 No.1