|Title:||A new strategy for intracellular delivery of enzyme using mesoporous silica nanoparticles: Superoxide dismutase||Authors:||Chen, Y.-P.
|Issue Date:||2013||Journal Volume:||135||Journal Issue:||4||Start page/Pages:||1516-1523||Source:||Journal of the American Chemical Society||Abstract:||
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. ? 2013 American Chemical Society.
|DOI:||10.1021/ja3105208||metadata.dc.subject.other:||Antioxidant enzyme; Cell-penetrating peptide; Cu ,Zn superoxide dismutase; E. coli; Enzyme delivery; Expression vectors; Flow cytometry analysis; Functional recovery; Fusion proteins; Human immunodeficiency virus-1; Intracellular delivery; Mesoporous silica nanoparticles; Molecular therapy; Oxidative damage; Reactive oxygen species; Super oxide dismutase; Superoxide dismutases; Therapeutic delivery; Transactivators; Transmembranes; Cell death; Confocal microscopy; Diseases; Escherichia coli; Free radicals; Gene expression; Oxygen; Purification; Viruses; Zinc; Enzymes; hybrid protein; mesoporous silica nanoparticle; nanoparticle; superoxide dismutase; transactivator protein; unclassified drug; apoptosis; article; confocal microscopy; conjugation; Escherichia coli; flow cytometry; protein expression; Apoptosis; Cell Membrane; Drug Delivery Systems; Flow Cytometry; HeLa Cells; Humans; Microscopy, Confocal; Nanoparticles; Particle Size; Porosity; Reactive Oxygen Species; rev Gene Products, Human Immunodeficiency Virus; Silicon Dioxide; Superoxide Dismutase; Surface Properties
|Appears in Collections:||化學系|
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