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http://hdl.handle.net/2445/177036
Title: | Biological properties of poly-L-lysine/DNA complexes generated by cooperative binding of the polycation |
Author: | Liu, Ge Molas, Maria Grossmann, Gregory A. Pasumarthy, Murali Perales Losa, Carlos Cooper, Mark J. Hanson, Richard W. |
Keywords: | ADN Metabolisme Lisina DNA Metabolism Lysine |
Issue Date: | 14-Sep-2001 |
Publisher: | American Society for Biochemistry and Molecular Biology |
Abstract: | We have evaluated the effect of NaCl concentration on the mode of binding of poly-L-lysine to DNA and the resulting structural and functional features of the condensed DNA particles using DNA precipitation, DNase I resistance, electron microscopy, and receptor-mediated gene transfer assays. At a high concentration of NaCl and in the presence of excess DNA, poly-L-lysine interacted with DNA cooperatively, fully condensing some of the DNA and leaving the rest of the DNA unbound. At low NaCl concentrations, poly-L-lysine molecules interacted with DNA in a noncooperative fashion, i.e. they bind randomly to the whole population of DNA molecules. Cooperative binding of poly-L-lysine to DNA occurred over a narrow range of NaCl concentrations, and the specific salt concentration depended on the length of the poly-L-lysine. The ability of condensed DNA to withstand digestion by DNase I was correlated with the structural features of the condensed DNA as determined by electron microscopy. Using our condensation procedure, cooperative binding of poly-L-lysine to DNA is a necessary prerequisite for the preparation of condensed DNA having a spherical shape and a diameter of 15-30 nm. Condensed DNA, containing galactosylated poly-L-lysine, was evaluated further for the extent and specificity of receptor-mediated gene transfer into HuH-7 human hepatoma cells via the asialoglycoprotein receptor. Efficient receptor-mediated transfection occurred only when condensed DNA complexes had a spherical shape with a diameter of 15-30 nm; asialofetuin, a natural ligand for the asialoglycoprotein receptor, inhibited this process by up to 90%. Our results support the importance of appropriate DNA condensation for the uptake and ultimate expression of DNA in hepatic cells. |
Note: | Reproducció del document publicat a: https://doi.org/10.1074/jbc.M105250200 |
It is part of: | Journal of Biological Chemistry, 2001, vol. 276, num. 37, p. 34379-34387 |
URI: | http://hdl.handle.net/2445/177036 |
Related resource: | https://doi.org/10.1074/jbc.M105250200 |
ISSN: | 0021-9258 |
Appears in Collections: | Articles publicats en revistes (Ciències Fisiològiques) |
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File | Description | Size | Format | |
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174985.pdf | 1.42 MB | Adobe PDF | View/Open |
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