title: Differential endosomal pathways for radically modified peptide vectors.
abstract: In the current work we characterize the uptake mechanism of two NickFect family members, NF51 and NF1, related to the biological activity of transfected plasmid DNA (pDNA). Both vectors condense pDNA into small negatively charged nanoparticles that transfect HeLa cells with equally high efficacy and the delivery is mediated by SCARA3 and SCARA5 receptors. NF1 condenses DNA into less homogeneous and less stable nanoparticles than NF51. NF51/pDNA nanoparticles enter the cells via macropinocytosis, while NF1/pDNA complexes use clathrin- or caveolae-mediated endocytosis and macropinocytosis. Analysis of separated endosomal compartments uncovered lysomotropic properties of NF51 that was also proven by cotransfection with chloroquine. In summary we characterize how radical modifications in peptides, such as introducing a kink in the structure of NF51 or including extra negative charge by phospho-tyrosine substitution in NF1, resulted in equally high efficacy for gene delivery, although this efficacy is achieved by using differential transfection pathways.
authors: Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu , Nooruse 1, 50411 Tartu, Estonia.
authors: Piret
authors: P
authors: Arukuusk
authors: Ly
authors: L
authors: Pärnaste
authors: Helerin
authors: H
authors: Margus
authors: N K Jonas
authors: NK
authors: Eriksson
authors: Luis
authors: L
authors: Vasconcelos
authors: Kärt
authors: K
authors: Padari
authors: Margus
authors: M
authors: Pooga
authors: Ulo
authors: U
authors: Langel
mesh_terms: D002966:Clathrin / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D014162:Transfection
chemical_list: D002966:Clathrin; D010455:Peptides; D004247:DNA