Transduced PEP-1-Grb7 fusion protein suppressed LPS-induced COX-2 expression.
Although the incidence and severity of atopic dermatitis (AD) is steadily increasing at an alarming rate, its pathogenic mechanisms remain poorly understood yet. Recently, we found that the expression of Grb7 protein was markedly decreased in AD patients using proteomic analysis. In the present study, human Grb7 gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-Grb7 fusion protein. The expressed and purified PEP-1-Grb7 fusion proteins transduced efficiently into skin cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced PEP-1-Grb7 protein was stable for 48 h. In addition, transduced PEP-1-Grb7 fusion protein markedly increased cell viability in macrophage RAW 264.7 cells treated with LPS by inhibition of the COX-2 expression level. These results suggest that the PEP-1-Grb7 fusion protein can be used in protein therapy for inflammatory skin disorders, including AD.
An
Jae Jin
JJ
Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
Kim
So Young
SY
Lee
Sun Hwa
SH
Kim
Dae Won
DW
Ryu
Hea Jin
HJ
Yeo
Seung Il
SI
Jang
Sang Ho
SH
Kwon
Hyung Joo
HJ
Kim
Tae Yoon
TY
Lee
Sang Chul
SC
Poo
Haryoung
H
Cho
Sung-Woo
SW
Lee
Kil Soo
KS
Park
Jinseu
J
Eum
Won Sik
WS
Choi
Soo Young
SY
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000378:metabolism; D015536:Down-Regulation; D005347:Fibroblasts / Q000187:drug effects; D051382:GRB7 Adaptor Protein / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014161:Transduction, Genetic
D008070:Lipopolysaccharides; C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D051382:GRB7 Adaptor Protein; D003543:Cysteamine; D051546:Cyclooxygenase 2