In vivo protein transduction: biologically active intact pep-1-superoxide dismutase fusion protein efficiently protects against ischemic insult.
Reactive oxygen species (ROS) are implicated in reperfusion injury after transient focal cerebral ischemia. The antioxidant enzyme Cu,Zn-superoxide dismutase (SOD) is one of the major means by which cells counteract the deleterious effects of ROS after ischemia. Recently, we reported that denatured Tat-SOD fusion protein is transduced into cells and skin tissue. Moreover, PEP-1 peptide, which has 21 amino acid residues, is a known carrier peptide that delivers full-length native proteins in vitro and in vivo. In the present study, we investigated the protective effects of PEP-1-SOD fusion protein after ischemic insult. A human SOD gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-SOD fusion protein. The expressed and purified fusion proteins were efficiently transduced both in vitro and in vivo with a native protein structure. Immunohistochemical analysis revealed that PEP-1-SOD injected intraperitoneally (i.p.) into mice can have access into brain neurons. When i.p.-injected into gerbils, PEP-1-SOD fusion proteins prevented neuronal cell death in the hippocampus caused by transient forebrain ischemia. These results suggest that the biologically active intact forms of PEP-1-SOD provide a more efficient strategy for therapeutic delivery in various human diseases related to this antioxidant enzyme or to ROS, including stroke.
Eum
Won Sik
WS
Department of Genetic Engineering, Research Institute for Bioscience and Biotechnology, Hallym University, 1-1 Okchon-Dong, Chunchon, Kangwon-Do, Korea, 200-702, Korea.
Kim
Dae Won
DW
Hwang
In Koo
IK
Yoo
Ki-Yeon
KY
Kang
Tae-Cheon
TC
Jang
Sang Ho
SH
Choi
Hee Soon
HS
Choi
Soo Hyun
SH
Kim
Young Hoon
YH
Kim
So Young
SY
Kwon
Hyeok Yil
HY
Kang
Jung Hoon
JH
Kwon
Oh-Shin
OS
Cho
Sung-Woo
SW
Lee
Kil Soo
KS
Park
Jinseu
J
Won
Moo Ho
MH
Choi
Soo Young
SY
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000201:enzymology; D002545:Brain Ischemia / Q000201:enzymology / Q000473:pathology; D003001:Cloning, Molecular; D000078404:Epidermal Cells; D004817:Epidermis / Q000201:enzymology*; D005849:Gerbillinae; D006624:Hippocampus / Q000201:enzymology* / Q000473:pathology; D008297:Male; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000201:enzymology; D010455:Peptides / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism*
D010455:Peptides; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase