Transduced PEP-1-AMPK inhibits the LPS-induced expression of COX-2 and iNOS in Raw264.7 cells.
AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme that plays a central role in cellular metabolic stress. Modulation of nitric oxide (NO) and cyclooxygenase-2 (COX-2) is considered a promising approach for the treatment of inflammation and neuronal diseases. In this study, the AMPK gene was fused in-frame with PEP-1 peptide in a bacterial expression vector to produce a PEP-1-AMPK fusion protein. Expressed and purified PEP-1-AMPK fusion proteins were transduced efficiently into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced PEP-1-AMPK fusion protein markedly inhibited LPS-induced iNOS and COX-2 expression. These results suggest that the PEP-1-AMPK fusion protein can be used for the protein therapy of COX-2 and NO-related disorders such as inflammation and neuronal diseases. [BMB reports 2010; 43(1): 40-45].
Shin
Min Jea
MJ
Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University Medical Center, Chunchon 200-704, Korea.
Lee
Yeom Pyo
YP
Kim
Dae Won
DW
An
Jae Jin
JJ
Jang
Sang Ho
SH
Cho
Sung-Min
SM
Sheen
Seung-Hoon
SH
Lee
Hae-Ran
HR
Kweon
Hae Yong
HY
Kang
Seok-Woo
SW
Lee
Kwang-Gill
KG
Park
Jinseu
J
Eum
Won Sik
WS
Cho
Yong-Jun
YJ
Choi
Soo Young
SY
D055372:AMP-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification; D013997:Time Factors; D014161:Transduction, Genetic
D008070:Lipopolysaccharides; C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D003543:Cysteamine; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D051546:Cyclooxygenase 2; D055372:AMP-Activated Protein Kinases