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http://hdl.handle.net/2445/177036
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DC Field | Value | Language |
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dc.contributor.author | Liu, Ge | - |
dc.contributor.author | Molas, Maria | - |
dc.contributor.author | Grossmann, Gregory A. | - |
dc.contributor.author | Pasumarthy, Murali | - |
dc.contributor.author | Perales Losa, Carlos | - |
dc.contributor.author | Cooper, Mark J. | - |
dc.contributor.author | Hanson, Richard W. | - |
dc.date.accessioned | 2021-05-05T15:16:55Z | - |
dc.date.available | 2021-05-05T15:16:55Z | - |
dc.date.issued | 2001-09-14 | - |
dc.identifier.issn | 0021-9258 | - |
dc.identifier.uri | http://hdl.handle.net/2445/177036 | - |
dc.description.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. | - |
dc.format.extent | 9 p. | - |
dc.format.mimetype | application/pdf | - |
dc.language.iso | eng | - |
dc.publisher | American Society for Biochemistry and Molecular Biology | - |
dc.relation.isformatof | Reproducció del document publicat a: https://doi.org/10.1074/jbc.M105250200 | - |
dc.relation.ispartof | Journal of Biological Chemistry, 2001, vol. 276, num. 37, p. 34379-34387 | - |
dc.relation.uri | https://doi.org/10.1074/jbc.M105250200 | - |
dc.rights | (c) American Society for Biochemistry and Molecular Biology, 2001 | - |
dc.source | Articles publicats en revistes (Ciències Fisiològiques) | - |
dc.subject.classification | ADN | - |
dc.subject.classification | Metabolisme | - |
dc.subject.classification | Lisina | - |
dc.subject.other | DNA | - |
dc.subject.other | Metabolism | - |
dc.subject.other | Lysine | - |
dc.title | Biological properties of poly-L-lysine/DNA complexes generated by cooperative binding of the polycation | - |
dc.type | info:eu-repo/semantics/article | - |
dc.type | info:eu-repo/semantics/publishedVersion | - |
dc.identifier.idgrec | 174985 | - |
dc.date.updated | 2021-05-05T15:16:56Z | - |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | - |
dc.identifier.pmid | 11438546 | - |
Appears in Collections: | Articles publicats en revistes (Ciències Fisiològiques) |
Files in This Item:
File | Description | Size | Format | |
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174985.pdf | 1.42 MB | Adobe PDF | View/Open |
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