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Inhibition of LPS-induced nitric oxide production by transduced Tat-arginine deiminase fusion protein in Raw 264.7 cells

  • Lee, Min-Jung (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Kim, Dae-Won (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Lee, Yeom-Pyo (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Jeong, Hoon-Jae (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Kang, Hye-Won (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Shin, Min-Jae (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Sohn, Eun-Jeong (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Kim, Mi-Jin (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Jang, Sang-Ho (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Kang, Tae-Cheon (Department of Anatomy and Neurobiology, College of Medicine, Hallym University) ;
  • Won, Moo-Ho (Department of Anatomy and Neurobiology, College of Medicine, Hallym University) ;
  • Min, Bon-Hong (Department of Pharmacology, College of Medicine, Korea University) ;
  • Cho, Sung-Woo (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine) ;
  • Lee, Kil-Soo (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Park, Jin-Seu (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University) ;
  • Eum, Won-Sik (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology) ;
  • Choi, Soo-Young (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology)
  • Published : 2009.05.31

Abstract

Arginine deiminase (ADI), an arginine-degrading enzyme, has anti-proliferative and anti-tumor activities and is capable of inhibiting the production of nitric oxide (NO). Modulation of nitric oxide (NO) production is considered a promising approach for the treatment of various diseases including cancer, inflammation and neuronal disorders. In this study, an ADI gene was fused with an HIV-1 Tat peptide in a bacterial expression vector to produce an genetic in-frame Tat-ADI fusion protein. When added exogenously to the culture media, the expressed and purified Tat-ADI fusion proteins were efficiently transduced into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced Tat-ADI fusion proteins markedly increased cell viability in cells treated with lipopolysaccharide (LPS). This increase in viability was mediated by an inhibition of NO production. These results suggest that this Tat-ADI fusion protein can be used in protein therapies of NO-related disorders such as cancer, inflammation and neuronal diseases.

Keywords

Arginine deiminase (ADI);HIV-1 Tat peptide;Inflammation;Nitric oxide synthase;Protein transduction

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