title: Protein Delivery System Containing a Nickel-Immobilized Polymer for Multimerization of Affinity-Purified His-Tagged Proteins Enhances Cytosolic Transfer.
abstract: Recombinant proteins with cytosolic or nuclear activities are emerging as tools for interfering with cellular functions. Because such tools rely on vehicles for crossing the plasma membrane we developed a protein delivery system consisting in the assembly of pyridylthiourea-grafted polyethylenimine (πPEI) with affinity-purified His-tagged proteins pre-organized onto a nickel-immobilized polymeric guide. The guide was prepared by functionalization of an ornithine polymer with nitrilotriacetic acid groups and shown to bind several His-tagged proteins. Superstructures were visualized by electron and atomic force microscopy using 2 nm His-tagged gold nanoparticles as probes. The whole system efficiently carried the green fluorescent protein, single-chain antibodies or caspase 3, into the cytosol of living cells. Transduction of the protease caspase 3 induced apoptosis in two cancer cell lines, demonstrating that this new protein delivery method could be used to interfere with cellular functions.
authors.affiliation: UMR 7199 and UMR 7213 CNRS Université de Strasbourg, Faculté de Pharmacie, Illkirch (France).
authors.forename: Viktoriia
authors.initials: V
authors.lastname: Postupalenko
authors.affiliation: UMR 7242, Ecole supérieure de biotechnologie Strasbourg, Illkirch (France).
authors.forename: Dominique
authors.initials: D
authors.lastname: Desplancq
authors.affiliation: Centre for Integrative Biology (CBI), IGBMC, Illkirch (France).
authors.forename: Igor
authors.initials: I
authors.lastname: Orlov
authors.affiliation: UMR 7199 and UMR 7213 CNRS Université de Strasbourg, Faculté de Pharmacie, Illkirch (France).
authors.forename: Youri
authors.initials: Y
authors.lastname: Arntz
authors.affiliation: Centre for Integrative Biology (CBI), IGBMC, Illkirch (France).
authors.forename: Danièle
authors.initials: D
authors.lastname: Spehner
authors.affiliation: UMR 7199 and UMR 7213 CNRS Université de Strasbourg, Faculté de Pharmacie, Illkirch (France).
authors.forename: Yves
authors.initials: Y
authors.lastname: Mely
authors.affiliation: Centre for Integrative Biology (CBI), IGBMC, Illkirch (France).
authors.forename: Bruno P
authors.initials: BP
authors.lastname: Klaholz
authors.affiliation: Centre for Integrative Biology (CBI), IGBMC, Illkirch (France).
authors.forename: Patrick
authors.initials: P
authors.lastname: Schultz
authors.affiliation: UMR 7242, Ecole supérieure de biotechnologie Strasbourg, Illkirch (France).
authors.forename: Etienne
authors.initials: E
authors.lastname: Weiss
authors.affiliation: UMR 7199 and UMR 7213 CNRS Université de Strasbourg, Faculté de Pharmacie, Illkirch (France). zuber@unistra.fr.
authors.forename: Guy
authors.initials: G
authors.lastname: Zuber
mesh_terms: D000345:Affinity Labels; D020285:Cryoelectron Microscopy; D003600:Cytosol / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D009532:Nickel / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000008:administration & dosage*
chemical_list: D000345:Affinity Labels; D011108:Polymers; D011506:Proteins; D006639:Histidine; D009532:Nickel
keywords: drug delivery; nanocarriers; protein delivery; proteins; self-assembly