Enhanced transduction of Cu,Zn-superoxide dismutase with HIV-1 Tat protein transduction domains at both termini.
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.
Eum
Won Sik
WS
Department of Genetic Engineering and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
Jang
Sang Ho
SH
Kim
Dae Won
DW
Choi
Hee Soon
HS
Choi
Soo Hyun
SH
Kim
So Young
SY
An
Jae Jin
JJ
Lee
Sun Hwa
SH
Han
Kyuhyung
K
Kang
Jung Hoon
JH
Kang
Tae-Cheon
TC
Won
Moo Ho
MH
Cho
Yong Joon
YJ
Choi
Jin Hi
JH
Kim
Tae Yoon
TY
Park
Jinseu
J
Choi
Soo Young
SY
D000818:Animals; D002470:Cell Survival; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015696:Gene Products, tat / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010957:Plasmids; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
D015696:Gene Products, tat; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase; D001616:beta-Galactosidase