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Transduction of Cu,Zn‐superoxide dismutase mediated by an HIV‐1 Tat protein basic domain into mammalian cells
Author(s) -
Kwon Hyeok Yil,
Eum Won Sik,
Jang Hyun Woo,
Kang Jung Hoon,
Ryu Jiyoon,
Ryong Lee Byung,
Jin Li Hua,
Park Jinseu,
Choi Soo Young
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)02215-8
Subject(s) - fusion protein , superoxide dismutase , transduction (biophysics) , microbiology and biotechnology , transactivation , hela , escherichia coli , biology , biochemistry , enzyme , chemistry , signal transduction , cell culture , gene expression , gene , cell , recombinant dna , genetics
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.