title: Enhanced transduction of Cu,Zn-superoxide dismutase with HIV-1 Tat protein transduction domains at both termini.
abstract: 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.
authors.affiliation: Department of Genetic Engineering and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Eum
authors.forename: Sang Ho
authors.initials: SH
authors.lastname: Jang
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Hee Soon
authors.initials: HS
authors.lastname: Choi
authors.forename: Soo Hyun
authors.initials: SH
authors.lastname: Choi
authors.forename: So Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Jae Jin
authors.initials: JJ
authors.lastname: An
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Lee
authors.forename: Kyuhyung
authors.initials: K
authors.lastname: Han
authors.forename: Jung Hoon
authors.initials: JH
authors.lastname: Kang
authors.forename: Tae-Cheon
authors.initials: TC
authors.lastname: Kang
authors.forename: Moo Ho
authors.initials: MH
authors.lastname: Won
authors.forename: Yong Joon
authors.initials: YJ
authors.lastname: Cho
authors.forename: Jin Hi
authors.initials: JH
authors.lastname: Choi
authors.forename: Tae Yoon
authors.initials: TY
authors.lastname: Kim
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: 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
chemical_list: D015696:Gene Products, tat; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase; D001616:beta-Galactosidase