NDLI: TAT-Mediated Protein Transduction of Nogo Extracellular Peptide 1-40 and its Biological Activity

Content Provider	Springer Nature Link
Author	Wang, Qiang Gou, Xingchun Jin, Weilin Xiong, Lize Hou, Lichao Chen, Shaoyang Zhang, Hui Zhu, Xiaoling Xu, Lixian
Copyright Year	2008
Abstract	Aim Nogo extracellular peptide 1-40 (NEP1-40), a Nogo-66 antagonistic peptide, is one of the potential candidates for therapeutic intervention after central nervous system injury. This study is focused on the generation of TAT-NEP1-40 fusion protein and its transducible effects and biological activity. Methods TAT-NEP1-40 fusion protein was expressed in vitro. Transducible effects of TAT-NEP1-40 were analyzed by using immunofluorescence staining or Western blot in vitro and in vivo. The biological activity of TAT-NEP1-40 was assessed by its effects against oxygen and glucose deprivation (OGD)-induced PC12 cell damages. Results Our results showed that the TAT-NEP1-40 fusion protein was successfully expressed, purified, and refolded. Western blot analysis and immunofluorescence staining confirmed the delivery of TAT-NEP1-40 protein into PC12 cells and rat brains. OGD caused cell apoptosis or death, decreased cell viability, increased lactate dehydrogenase release in medium and the Bax/Bcl-2 ratio, all of which were prevented by the TAT-NEP1-40 fusion proteins when added exogenously to culture medium. In addition, TAT-NEP1-40 promoted neurite outgrowth of PC12 cells exposed to OGD. Conclusion These results demonstrate that the TAT-NEP1-40 can be successfully generated and efficiently transduced into PC12 cells and rat brains. The TAT-NEP1-40 can protect PC12 cells against OGD and promote neurite outgrowth. This finding suggests that the transducible TAT-NEP1-40 fusion protein offers a possibility of the development of novel therapy for cerebral injuries via delivery of the biologically active TAT-NEP1-40 fusion protein into injured sites.
Starting Page	97
Ending Page	108
Page Count	12
File Format	PDF
ISSN	02724340
Journal	Cellular and Molecular Neurobiology
Volume Number	29
Issue Number	1
e-ISSN	15736830
Language	English
Publisher	Springer US
Publisher Date	2008-08-29
Publisher Place	Boston
Access Restriction	Subscribed
Subject Keyword	Nogo NEP1-40 TAT Oxygen and glucose deprivation PC12 cells Animal Anatomy / Morphology / Histology Neurosciences
Content Type	Text
Resource Type	Article
Subject	Cell Biology Medicine Cellular and Molecular Neuroscience

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