Protein transduction of an antioxidant enzyme: subcellular localization of superoxide dismutase fusion protein in cells.
In protein therapy, it is important for exogenous protein to be delivered into the target subcellular localization. To transduce a therapeutic protein into its specific subcellular localization, we synthesized nuclear localization signal (NLS) and membrane translocation sequence signal (MTS) peptides and produced a genetic in-frame SOD fusion protein. The purified SOD fusion proteins were efficiently transduced into mammalian cells with enzymatic activities. Immunofluorescence and Western blot analysis revealed that the SOD fusion proteins successfully transduced into the nucleus and the cytosol in the cells. The viability of cells treated with paraquat was markedly increased by the transduced fusion proteins. Thus, our results suggest that these peptides should be useful for targeting the specific localization of therapeutic proteins in various human diseases.
Kim
Dae Won
DW
Department of Biomedical Science and the Research Institute for Bioscience and Biotechnology, Hallym University, Chuncheon, Korea.
Kim
So Young
SY
Lee
Sun Hwa
SH
Lee
Yeum Pyo
YP
Lee
Min Jung
MJ
Jeong
Min Seop
MS
Jang
Sang Ho
SH
Park
Jinseu
J
Lee
Kil Soo
KS
Kang
Tae-Cheon
TC
Won
Moo Ho
MH
Cho
Sung-Woo
SW
Kwon
Oh-Shin
OS
Eum
Won Sik
WS
Choi
Soo Young
SY
D000975:Antioxidants / Q000378:metabolism*; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003600:Cytosol / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006540:Herbicides / Q000494:pharmacology; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D019913:Nuclear Localization Signals / Q000235:genetics* / Q000378:metabolism; D018384:Oxidative Stress; D010269:Paraquat / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013347:Subcellular Fractions; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic
D000975:Antioxidants; D006540:Herbicides; D019913:Nuclear Localization Signals; D010446:Peptide Fragments; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase; D010269:Paraquat