Inhibition of LPS-induced nitric oxide production by transduced Tat-arginine deiminase fusion protein in Raw 264.7 cells.
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.
Lee
Min Jung
MJ
Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University, Chunchon 200-702, Korea.
Kim
Dae Won
DW
Lee
Yeom Pyo
YP
Jeong
Hoon Jae
HJ
Kang
Hye Won
HW
Shin
Min Jae
MJ
Sohn
Eun Jeong
EJ
Kim
Mi Jin
MJ
Jang
Sang Ho
SH
Kang
Tae-Cheon
TC
Won
Moo Ho
MH
Min
Bon-Hong
BH
Cho
Sung-Woo
SW
Lee
Kil Soo
KS
Park
Jinseu
J
Eum
Won Sik
WS
Choi
Soo Young
SY
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
D015696:Gene Products, tat; D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D006867:Hydrolases; C019723:arginine deiminase