title: Efficient delivery of p53 and cytochrome c by supramolecular assembly of a dendritic multi-domain delivery system.
abstract: Versatile nanocarrier systems facilitating uptake of exogenous proteins are highly alluring in evaluating these proteins for therapeutic applications. The self-assembly of an efficient nano-sized protein transporter consisting of three different entities is presented: A streptavidin protein core functioning as an adapter, second generation polyamidoamine dendrons for facilitating cell uptake as well as two different therapeutic proteins (tumor suppressor p53 or pro-apoptotic cytochrome c as cargo). Well-defined dendrons containing a biotin core are prepared and display no cytotoxic behavior upon conjugation to streptavidin. The integration of biotinylated human recombinant p53 (B-p53) into the three component system allows excellent internalization into HeLa, A549 and SaOS osteosarcoma cells monitored via confocal microscopy, immunoblot analysis and co-localization studies. In addition, the conjugation of B-p53 to dendronized streptavidin preserves its specific DNA-binding in vitro, and its delivery into SaOS cells impairs cell viability with concomitant activation of caspases 3 and 7. The versatility of this system is further exhibited by the significant enhancement of the pro-apoptotic effects of internalized cytochrome c which is analyzed by flow cytometry and cell viability assays. These results demonstrate that the "bio-click" self-assembly of biotinylated dendrons and proteins on a streptavidin adapter yields a stable supramolecular complex. This efficient bionanotransporter provides an attractive platform for mediating the delivery of functional proteins of interest into living mammalian cells in a facile and rapid way.
authors: Institute of Organic Chemistry III, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany; Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
authors: David Yuen Wah
authors: DY
authors: Ng
authors: Jörg
authors: J
authors: Fahrer
authors: Yuzhou
authors: Y
authors: Wu
authors: Klaus
authors: K
authors: Eisele
authors: Seah Ling
authors: SL
authors: Kuan
authors: Holger
authors: H
authors: Barth
authors: Tanja
authors: T
authors: Weil
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001710:Biotin / Q000737:chemistry; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D050091:Dendrimers / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D006367:HeLa Cells; D006801:Humans; D019809:Streptavidin / Q000737:chemistry; D016159:Tumor Suppressor Protein p53 / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics
chemical_list: D050091:Dendrimers; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D001710:Biotin; D045304:Cytochromes c; D019809:Streptavidin
keywords: biohybrids; dendronized proteins; p53 and cytochrome c delivery; streptavidin adapters; supramolecular drugs