Transduction of Cu,Zn‐superoxide dismutase mediated by an HIV‐1 Tat protein basic domain into mammalian cells

FEBS Letters

Eum, Won Sik¹  Jin, Li Hua³  Choi, Soo Young³  Kang, Jung Hoon²  Park, Jinseu³  Jang, Hyun Woo¹  Ryu, Jiyoon³  Ryong Lee, Byung³  Kwon, Hyeok Yil¹ 

[1] Department of Physiology, College of Medicine, Hallym University, Chunchon 200-702, South Korea;Department of Genetic Engineering, Division of Natural Sciences, Chongju University, Chongju 360-764, South Korea;Department of Genetic Engineering, Division of Life Sciences, Hallym University, Chunchon 200-702, South Korea

关键词: Transduction;    Copper;    zinc-superoxide dismutase;    Human immunodeficiency virus type 1 transactivator of transcription;    HIV-1;    human immunodeficiency virus type 1;    Tat;    transactivator of transcription;    Cu;    Zn-SOD;    Cu;    Zn-superoxide dismutase;    PCR;    polymerase chain reaction;    IPTG;    isopropyl-β-D-thiogalactoside;    SDS–PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    PBS;    phosphate-buffered saline;   

DOI  :  10.1016/S0014-5793(00)02215-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A human Cu,Zn-superoxide dismutase (Cu,Zn-SOD) gene was fused with a gene fragment encoding the nine amino acid transactivator of transcription (Tat) protein transduction domain (RKKRRQRRR) of HIV-1 in a bacterial expression vector to produce a genetic in-frame Tat–SOD fusion protein. The expressed and purified Tat–SOD fusion protein in Escherichia coli can enter HeLa cells in a time- and dose-dependent manner when added exogenously in a culture media. Denatured Tat–SOD protein was transduced much more efficiently into cells than were native proteins. Once inside the cells, transduced Tat–SOD protein was enzymatically active and stable for 24 h. The cell viability of HeLa cells treated with paraquat, an intracellular superoxide anion generator, was increased by transduced Tat–SOD. These lines of results suggest that the transduction of Tat–SOD fusion protein may be one of the ways to replenish the Cu,Zn-SOD in the various disorders related to this antioxidant enzyme.

【 授权许可】

Unknown