A new strategy for intracellular delivery of enzyme using mesoporous silica nanoparticles: Superoxide dismutase

Yi Ping Chen, Chien Tsu Chen, Yann Hung, Chih Ming Chou, Tsang Pai Liu, Ming Ren Liang, Chao Tsen Chen, Chung Yuan Mou

研究成果: 雜誌貢獻文章同行評審

118 引文 斯高帕斯(Scopus)

摘要

We developed mesoporous silica nanoparticle (MSN) as a multifunctional vehicle for enzyme delivery. Enhanced transmembrane delivery of a superoxide dismutase (SOD) enzyme embedded in MSN was demonstrated. Conjugation of the cell-penetrating peptide derived from the human immunodeficiency virus 1 (HIV) transactivator protein (TAT) to mesoporous silica nanoparticle is shown to be an effective way to enhance transmembrane delivery of nanoparticles for intracellular and molecular therapy. Cu,Zn-superoxide dismutase (SOD) is a key antioxidant enzyme that detoxifies intracellular reactive oxygen species, ROS, thereby protecting cells from oxidative damage. In this study, we fused a human Cu,Zn-SOD gene with TAT in a bacterial expression vector to produce a genetic in-frame His-tagged TAT-SOD fusion protein. The His-tagged TAT-SOD fusion protein was expressed in E. coli using IPTG induction and purified using FMSN-Ni-NTA. The purified TAT-SOD was conjugated to FITC-MSN forming FMSN-TAT-SOD. The effectiveness of FMSN-TAT-SOD as an agent against ROS was investigated, which included the level of ROS and apoptosis after free radicals induction and functional recovery after ROS damage. Confocal microscopy on live unfixed cells and flow cytometry analysis showed characteristic nonendosomal distribution of FMSN-TAT-SOD. Results suggested that FMSN-TAT-SOD may provide a strategy for the therapeutic delivery of antioxidant enzymes that protect cells from ROS damage.
原文英語
頁(從 - 到)1516-1523
頁數8
期刊Journal of the American Chemical Society
135
發行號4
DOIs
出版狀態已發佈 - 一月 30 2013

ASJC Scopus subject areas

  • 化學 (全部)
  • 催化
  • 生物化學
  • 膠體和表面化學

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