14 results on '"Eum, Won Sik"'
Search Results
2. Erratum to: “Amelioration of Streptozotocin-Induced Diabetes by Agrocybe chaxingu Polysaccharide”
- Author
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Lee, Byung Ryong, primary, Lee, Yeom Pyo, additional, Kim, Dae Won, additional, Song, Ha Yong, additional, Yoo, Ki-Yeon, additional, Won, Moo Ho, additional, Kang, Tae-Cheon, additional, Lee, Kwang Jae, additional, Kim, Kyung Hee, additional, Joo, Jin Ho, additional, Ham, Hun Ju, additional, Hur, Jang Hyun, additional, Cho, Sung-Woo, additional, Han, Kyu Hyung, additional, Lee, Kil Soo, additional, Park, Jinseu, additional, Eum, Won Sik, additional, and Choi, Soo Young, additional
- Published
- 2013
- Full Text
- View/download PDF
3. Transduced Tat-DJ-1 protein protects against oxidative stress-induced SH-SY5Y cell death and Parkinson disease in a mouse model
- Author
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Jeong, Hoon Jae, primary, Kim, Dae Won, additional, Woo, Su Jung, additional, Kim, Hye Ri, additional, Kim, So Mi, additional, Jo, Hyo Sang, additional, Park, Meeyoung, additional, Kim, Duk-Soo, additional, Kwon, Oh-Shin, additional, Hwang, In Koo, additional, Han, Kyu Hyung, additional, Park, Jinseu, additional, Eum, Won Sik, additional, and Choi, Soo Young, additional
- Published
- 2012
- Full Text
- View/download PDF
4. Amelioration of Streptozotocin-Induced Diabetes by Agrocybe chaxingu Polysaccharide
- Author
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Lee, Byung Ryong, primary, Lee, Yeom Pyo, additional, Kim, Dae Won, additional, Song, Ha Yong, additional, Yoo, Ki-Yeon, additional, Won, Moo Ho, additional, Kang, Tae-Cheon, additional, Lee, Kwang Jae, additional, Kim, Kyung Hee, additional, Joo, Jin Ho, additional, Ham, Hun Ju, additional, Hur, Jang Hyun, additional, Cho, Sung-Woo, additional, Han, Kyu Hyung, additional, Lee, Kil Soo, additional, Park, Jinseu, additional, Eum, Won Sik, additional, and Choi, Soo Young, additional
- Published
- 2010
- Full Text
- View/download PDF
5. Transduction of familial amyotrophic lateral sclerosis-related mutant PEP-1-SOD proteins into neuronal cells.
- Author
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An JJ, Lee YP, Kim SY, Lee SH, Kim DW, Lee MJ, Jeong MS, Jang SH, Kang JH, Kwon HY, Kang TC, Won MH, Cho SW, Kwon OS, Lee KS, Park J, Eum WS, and Choi SY
- Subjects
- Animals, Astrocytes cytology, Cell Survival, Cells, Cultured, Cysteamine metabolism, Heat-Shock Proteins genetics, Heat-Shock Proteins metabolism, Humans, Oxidative Stress, Peptides genetics, Rats, Rats, Wistar, Recombinant Fusion Proteins genetics, Superoxide Dismutase genetics, Superoxide Dismutase-1, Amyotrophic Lateral Sclerosis genetics, Amyotrophic Lateral Sclerosis metabolism, Astrocytes physiology, Cysteamine analogs & derivatives, Peptides metabolism, Recombinant Fusion Proteins metabolism, Superoxide Dismutase metabolism, Transduction, Genetic
- Abstract
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.
- Published
- 2008
6. Transduced human copper chaperone for Cu,Zn-SOD (PEP-1-CCS) protects against neuronal cell death.
- Author
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Choi SH, Kim DW, Kim SY, An JJ, Lee SH, Choi HS, Sohn EJ, Hwang SI, Won MH, Kang TC, Kwon HJ, Kang JH, Cho SW, Park J, Eum WS, and Choi SY
- Subjects
- Animals, Astrocytes metabolism, Brain Ischemia chemically induced, Brain Ischemia metabolism, Brain Ischemia pathology, Cell Death drug effects, Gerbillinae, Hippocampus drug effects, Humans, Immunoenzyme Techniques, Male, Neurons metabolism, Oxidative Stress physiology, Paraquat toxicity, Recombinant Fusion Proteins metabolism, Recombinant Fusion Proteins therapeutic use, Transfection, Copper metabolism, Molecular Chaperones therapeutic use, Neurons drug effects
- Abstract
Reactive oxygen species (ROS) contribute to the development of various human diseases. Cu,Zn-superoxide dismutase (SOD) is one of the major means by which cells counteract the deleterious effects of ROS. SOD activity is dependent upon bound copper ions supplied by its partner metallochaperone protein, copper chaperone for SOD (CCS). In the present study, we investigated the protective effects of PEP-1-CCS against neuronal cell death and ischemic insults. When PEP-1-CCS was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Moreover, transduced PEP-1-CCS markedly increased endogenous SOD activity in the cells. Immunohistochemical analysis revealed that it prevented neuronal cell death in the hippocampus in response to transient forebrain ischemia. These results suggest that CCS is essential to activate SOD, and that transduction of PEP-1-CCS provides a potential strategy for therapeutic delivery in various human diseases including stroke related to SOD or ROS.
- Published
- 2005
7. Enhanced transduction of Cu,Zn-superoxide dismutase with HIV-1 Tat protein transduction domains at both termini.
- Author
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Eum WS, Jang SH, Kim DW, Choi HS, Choi SH, Kim SY, An JJ, Lee SH, Han K, Kang JH, Kang TC, Won MH, Cho YJ, Choi JH, Kim TY, Park J, and Choi SY
- Subjects
- Animals, Cell Survival, Escherichia coli genetics, Escherichia coli metabolism, HeLa Cells, Humans, Mice, Plasmids, Protein Structure, Tertiary, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins isolation & purification, Recombinant Fusion Proteins metabolism, Skin cytology, Superoxide Dismutase genetics, beta-Galactosidase metabolism, Gene Products, tat metabolism, Skin metabolism, Superoxide Dismutase metabolism, Transduction, Genetic
- 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.
- Published
- 2005
8. Transduced Tat-SOD fusion protein protects against ischemic brain injury.
- Author
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Kim DW, Eum WS, Jang SH, Kim SY, Choi HS, Choi SH, An JJ, Lee SH, Lee KS, Han K, Kang TC, Won MH, Kang JH, Kwon OS, Cho SW, Kim TY, Park J, and Choi SY
- Subjects
- Animals, Antioxidants therapeutic use, Cell Death drug effects, Gene Products, tat therapeutic use, Gerbillinae, Male, Mice, Mice, Inbred ICR, Oxidative Stress physiology, Recombinant Fusion Proteins therapeutic use, Transduction, Genetic, Astrocytes cytology, Astrocytes drug effects, Ischemic Attack, Transient drug therapy, Reperfusion Injury prevention & control, Superoxide Dismutase therapeutic use
- Abstract
Reactive oxygen species (ROS) are implicated in reperfusion injury after transient focal cerebral ischemia. The antioxidant enzyme, Cu,Zn-superoxide dismutase (SOD), is one of the major means by which cells counteract the deleterious effects of ROS after ischemia. Recently, we reported that when Tat-SOD fusion protein is transduced into pancreatic beta cells it protects the beta cells from destruction by relieving oxidative stress in ROS-implicated diabetes (Eum et al., 2004). In the present study, we investigated the protective effects of Tat-SOD fusion protein against neuronal cell death and ischemic insults. When Tat-SOD was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Immunohistochemical analysis revealed that Tat-SOD injected intraperitoneally (i.p.) into mice has access to various tissues including brain neurons. When i.p. injected into gerbils, Tat-SOD prevented neuronal cell death in the hippocampus in response to transient fore-brain ischemia. These results suggest that Tat-SOD provides a strategy for therapeutic delivery in various hu-man diseases, including stroke, related to this anti-oxidant enzyme or to ROS.
- Published
- 2005
9. Site-directed mutagenesis of human brain GABA transaminase: lysine-357 is involved in cofactor binding at the active site.
- Author
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Kim DW, Yoon CS, Eum WS, Lee BR, An JJ, Lee SH, Lee SR, Ahn JY, Kwon OS, Kang TC, Won MH, Cho SW, Lee KS, Park J, and Choi SY
- Subjects
- Amino Acid Sequence, Base Sequence, Binding Sites, Catalysis, Electrophoresis, Polyacrylamide Gel, Genetic Vectors, Humans, Lysine chemistry, Molecular Sequence Data, Mutagenesis, Site-Directed, Mutation, Protein Binding, Pyridoxal Phosphate metabolism, Recombinant Proteins chemistry, Spectrophotometry, 4-Aminobutyrate Transaminase genetics, 4-Aminobutyrate Transaminase physiology, Brain enzymology, Lysine genetics
- Abstract
gamma-Aminobutyrate transaminase (GABA-T), a key enzyme of the GABA shunt, converts the major inhibitory neurotransmitter, GABA, to succinic semialdehyde. Although GABA-T is a pivotal factor implicated in the pathogenesis of various neurological disorders, its function remains to be elucidated. In an effort to clarify the structural and functional roles of specific lysyl residue in human brain GABA-T, we constructed human brain GABA-T mutants, in which the lysyl residue at position 357 was mutated to various amino acids including asparagine (K357N). The purified mutant GABA-T enzymes displayed neither catalytic activity nor absorption bands at 330 and 415 nm that are characteristic of pyridoxal-5'-phosphate (PLP) covalently linked to the protein. The wild type apoenzyme reconstituted with exogenous PLP had catalytic activity, while the mutant apoenzymes did not. These results indicate that lysine 357 is essential for catalytic function, and is involved in binding PLP at the active site.
- Published
- 2004
10. Cysteine-321 of human brain GABA transaminase is involved in intersubunit cross-linking.
- Author
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Yoon CS, Kim DW, Jang SH, Lee BR, Choi HS, Choi SH, Kim SY, An JJ, Kwon OS, Kang TC, Won MH, Cho SW, Lee KS, Park J, Eum WS, and Choi SY
- Subjects
- 4-Aminobutyrate Transaminase genetics, Brain, Catalysis, Cross-Linking Reagents, Disulfides, Dithionitrobenzoic Acid, Humans, Mutagenesis, Site-Directed, Protein Structure, Quaternary, Protein Subunits, 4-Aminobutyrate Transaminase chemistry, 4-Aminobutyrate Transaminase metabolism, Cysteine metabolism
- Abstract
Gamma-aminobutyrate transaminase (GABA-T), a key homodimeric enzyme of the GABA shunt, converts the major inhibitory neurotransmitter GABA to succinic semialdehyde. We previously overexpressed, purified and characterized human brain GABA-T. To identify the structural and functional roles of the cysteinyl residue at position 321, we constructed various GABA-T mutants by site-directed mutagenesis. The purified wild type GABA-T enzyme was enzymatically active, whereas the mutant enzymes were inactive. Reaction of 1.5 sulfhydryl groups per wild type dimer with 5,5 cent-dithiobis-2-nitrobenzoic acid (DTNB) produced about 95% loss of activity. No reactive -SH groups were detected in the mutant enzymes. Wild type GABA-T, but not the mutants, existed as an oligomeric species of Mr = 100,000 that was dissociable by 2-mercaptoethanol. These results suggest that the Cys321 residue is essential for the catalytic function of GABA-T, and that it is involved in the formation of a disulfide link between two monomers of human brain GABA-T.
- Published
- 2004
11. Ginsenosides enhance the transduction of tat-superoxide dismutase into mammalian cells and skin.
- Author
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Kim DW, Eum WS, Jang SH, Yoon CS, Choi HS, Choi SH, Kim YH, Kim SY, Lee ES, Baek NI, Kwon HY, Choi JH, Choi YC, Kwon OS, Cho SW, Han K, Lee KS, Park J, and Choi SY
- Subjects
- Animals, Gene Products, tat genetics, HeLa Cells, Humans, Immunohistochemistry, Mice, Protein Transport physiology, Recombinant Fusion Proteins metabolism, Skin metabolism, Superoxide Dismutase genetics, Gene Products, tat metabolism, Ginsenosides metabolism, Recombinant Fusion Proteins genetics, Superoxide Dismutase metabolism
- Abstract
We previously reported that Tat-Cu,Zn-superoxide dismutase (Tat-SOD), a major antioxidant enzyme, can be directly transduced into mammalian cells and skin [Kwon et al. (2000); Park et al. (2002)]. To enhance the therapeutic potential of Tat-SOD in the treatment of various disorders, we screened a number of natural products for their ability to increase transduction efficiency. Ginsenosides were effective with cultured HeLa cells and enhanced the penetration of Tat-SOD into both the epidermis and the dermis of the subcutaneous layer when sprayed on mice skin. Although their mechanism of action is not fully understood we believe that ginsenosides may be useful cofactors with this antioxidant enzyme in anti-aging cosmetics or as a therapeutic protein in disorders related to reactive-oxygen species.
- Published
- 2003
12. Human liver catalase: cloning, expression and characterization of monoclonal antibodies.
- Author
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Jin LH, Kim DW, Eum WS, Yoon CS, Jang SH, Choi HS, Choi SH, Kim YH, Kim SY, Jung MR, Kang TC, Won MH, Lee HY, Kang JH, Kwon OS, Cho SW, Lee KS, Park J, and Choi SY
- Subjects
- Amino Acid Sequence, Animals, Antibodies, Monoclonal biosynthesis, Catalase genetics, Catalase isolation & purification, Cell Line, Cloning, Molecular, Cross Reactions, Escherichia coli genetics, Gene Library, Humans, Mice, Molecular Sequence Data, Rats, Sequence Alignment, Antibodies, Monoclonal genetics, Catalase immunology, Liver enzymology
- Abstract
We isolated a cDNA encoding liver catalase from a human liver cDNA library. The cDNA had a high degree of sequence similarity to the corresponding enzyme from other sources. It was expressed in E. coli using the pET15b vector. The protein produced was enzymatically active after purification, and its kinetic parameters closely resembled those of other mammalian catalases. Monoclonal antibodies were generated against the purified catalase; six antibodies recognizing different epitopes were obtained, one of which inhibited the enzyme. The cross reactions of the antibodies with brain catalases from human and other mammalian tissues were investigated, and all the immunoreactive bands obtained on Western blots had molecular masses of about 58 kDa. Similarly fractionated extracts of several mammalian cell lines all gave a single band of molecular mass 58 kDa. These results indicate that mammalian livers and human cell lines contain only one major type of immunologically reactive catalase, even though some of catalases have been previously reported to differ in certain properties.
- Published
- 2003
13. 9-polylysine protein transduction domain: enhanced penetration efficiency of superoxide dismutase into mammalian cells and skin.
- Author
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Park J, Ryu J, Jin LH, Bahn JH, Kim JA, Yoon CS, Kim DW, Han KH, Eum WS, Kwon HY, Kang TC, Won MH, Kang JH, Cho SW, and Choi SY
- Subjects
- Animals, Antioxidants metabolism, Cell Survival, Cells, Cultured, Fibroblasts cytology, Fibroblasts drug effects, Fibroblasts metabolism, Gene Products, tat genetics, Humans, Immunohistochemistry, Mice, Paraquat pharmacology, Polylysine genetics, Protein Structure, Tertiary, Protein Transport, Reactive Oxygen Species metabolism, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Skin cytology, Superoxide Dismutase genetics, Free Radical Scavengers metabolism, Polylysine metabolism, Skin metabolism, Superoxide Dismutase metabolism, Transduction, Genetic
- Abstract
Antioxidant enzymes, such as superoxide dismutase (SOD) and catalase (CAT), have been considered to have a beneficial effect against various diseases that are mediated by the reactive oxygen species (ROS). Although a variety of modified recombinant antioxidant enzymes have been generated to protect against oxidative stresses, the lack of their transduction ability into cells resulted in a limited ability to detoxify intracellular ROS. To render the SOD enzyme capable of detoxifying intracellular ROS when added extracellularly, cell-permeable recombinant SOD proteins were generated. A human Cu,Zn-superoxide dismutase (Cu,Zn-SOD) gene was fused with a gene fragment that encodes the 9 amino acids Tat protein transduction domain (RKKRRQRRR) of HIV-1 and lysine rich peptide (KKKKKKKKK) in a bacterial expression vector in order to produce a genetic in-frame Tat-SOD and 9Lys-SOD fusion protein, respectively. The expressed and purified Tat-SOD and 9Lys-SOD fusion proteins can transduce into human fibroblast cells, and they were enzymatically active and stable for 24 h. The cell viability of the fibroblast cells that were treated with paraquat, an intracellular superoxide anion generator, was increased by the transduced Tat-SOD or 9Lys-SOD. The transduction efficacy of 9Lys-SOD was more efficient than that of Tat-SOD. We evaluated the ability of the SOD fusion pmteins to transduce into animal skin. This analysis showed that Tat-SOD and 9Lys-SOD fusion proteins efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin (judged by the immunohistochemistry and specific enzyme activities). The enzymatic activity of the transduced 9Lys-SOD was higher than that of Tat-SOD, indicating that the penetration of 9Lys-SOD was more efficient when put into the skin. These results suggest Tat-SOD and 9Lys-SOD fusion proteins can be used as anti-aging cosmetics, or in protein therapy, for various disorders that are related to this antioxidant enzyme and ROS.
- Published
- 2002
14. Transduction efficacy of Tat-Cu,Zn-superoxide dismutase is enhanced by copper ion recovery of the fusion protein.
- Author
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Eum WS, Choung IS, Kim AY, Lee YJ, Kang JH, Park J, Lee KS, Kwon HY, and Choi SY
- Subjects
- Antioxidants metabolism, Cell Survival, HeLa Cells, Humans, Metals metabolism, Paraquat metabolism, Protein Transport, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics, Superoxide Dismutase chemistry, Superoxide Dismutase metabolism, Copper metabolism, Gene Products, tat genetics, Recombinant Fusion Proteins metabolism, Superoxide Dismutase genetics
- Abstract
We previously reported that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. To enhance the therapeutic potential of Tat-SOD for the treatment of various disorders that are related to this antioxidant enzyme, the transduction efficacy of Tat-SOD should be heightened. Therefore, we investigated whether copper ion recovery of the fusion protein could enhance the transduction potential of Tat-SOD in cultured HeLa cells. The results showed that the transduction potential of Tat-SOD was markedly enhanced by copper ions, and moderately increased by zinc ions. Compared with Tat-SOD, the Tat-SOD that recovered the copper ion (CR-Tat-SOD) achieved a significant increase in intracellular concentration and enzymatic activity. Therefore, CR-Tat-SOD was transduced into HeLa cells in a rapid saturation manner, but Tat-SOD was shown in a time-dependent manner. With the higher transduction efficacy of CR-Tat-SOD than that of Tat-SOD, the transduced CR-Tat-SOD significantly increased the viability of HeLa cells that were pretreated with paraquat, an intracellular superoxide anion generator. Although the mechanism of the enhanced transduction of Tat-SOD by copper ions is still unanswered, these results indicate that copper ions facilitate the transduction of SOD. These then significantly increase the biological effectiveness of this antioxidant enzyme.
- Published
- 2002
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