title: TAT-mediated protein transduction of Nogo extracellular peptide 1-40 and its biological activity.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi'an, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wang
authors.forename: Xingchun
authors.initials: X
authors.lastname: Gou
authors.forename: Weilin
authors.initials: W
authors.lastname: Jin
authors.forename: Lize
authors.initials: L
authors.lastname: Xiong
authors.forename: Lichao
authors.initials: L
authors.lastname: Hou
authors.forename: Shaoyang
authors.initials: S
authors.lastname: Chen
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Zhu
authors.forename: Lixian
authors.initials: L
authors.lastname: Xu