title: Inhibition of LPS-induced nitric oxide production by transduced Tat-arginine deiminase fusion protein in Raw 264.7 cells.
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.
authors.affiliation: Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University, Chunchon 200-702, Korea.
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Lee
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Yeom Pyo
authors.initials: YP
authors.lastname: Lee
authors.forename: Hoon Jae
authors.initials: HJ
authors.lastname: Jeong
authors.forename: Hye Won
authors.initials: HW
authors.lastname: Kang
authors.forename: Min Jae
authors.initials: MJ
authors.lastname: Shin
authors.forename: Eun Jeong
authors.initials: EJ
authors.lastname: Sohn
authors.forename: Mi Jin
authors.initials: MJ
authors.lastname: Kim
authors.forename: Sang Ho
authors.initials: SH
authors.lastname: Jang
authors.forename: Tae-Cheon
authors.initials: TC
authors.lastname: Kang
authors.forename: Moo Ho
authors.initials: MH
authors.lastname: Won
authors.forename: Bon-Hong
authors.initials: BH
authors.lastname: Min
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Kil Soo
authors.initials: KS
authors.lastname: Lee
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Eum
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D015696:Gene Products, tat / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism; D006801:Humans; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis*; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D015696:Gene Products, tat; D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D006867:Hydrolases; C019723:arginine deiminase