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| - Surface Membrane Biotinylation Efficiently Mediates theEndocytosis of Avidin Bioconjugates into Nucleated Cells
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| - Here we demonstrate that biotin covalently attached to cell surface obligates existing receptors toendocytose avidin bioconjugates into nucleated cells. Incubation of fluorescein-labeled avidin withbiotinylated cell lines resulted in uniform and rapid surface attachment and endocytosis comparedwith no detectable association of the avidin-conjugated dye with unbiotinylated cells. Uptake wasdetected within minutes with efficiencies approaching 100% in cell lines and freshly obtained peripheralblood mononuclear cells. After 24 h, avidin was barely detectable on the surface of the nucleatedcells. In marked contrast, fluorescent avidin remained exclusively on the external membrane oferythrocytes after 24 h. To investigate biotin-mediated endocytosis for the delivery of DNA, we preparedpolyethylenimine−avidin (PEI−avidin) conjugates. Surface biotinylation significantly increased thetransfection efficiencies of PEI−avidin condensed plasmid DNA coding green fluorescent protein (GFP)to the level of transferrin-receptor targeted gene delivery (15−20% GFP positive cells in culture after48 h). The increase in transfection efficiency was blocked by the addition of free avidin or biotin tothe culture medium. Biotin covalently bound to cell surface membrane proteins efficiently mediatesthe entry of avidin bioconjugates into nucleated cells.
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