JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Belting, M.
Right arrow Articles by Petersson, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Belting, M.
Right arrow Articles by Petersson, P.

J Biol Chem, Vol. 274, Issue 27, 19375-19382, July 2, 1999

Intracellular Accumulation of Secreted Proteoglycans Inhibits Cationic Lipid-mediated Gene Transfer
CO-TRANSFER OF GLYCOSAMINOGLYCANS TO THE NUCLEUS

Mattias Belting and Per Petersson

From the Department of Cell and Molecular Biology, Section for Cell and Matrix Biology, Lund University, P. O. Box 94, S-221 00 Lund, Sweden

Molecules secreted by potential target cells may interfere with cationic lipid-mediated gene transfer. This has been studied using human lung fibroblasts and human epidermoid lung cancer cells. Secreted cell medium components caused a substantial decrease both in the uptake of cationic lipid-DNA complexes (2-4-fold) and in reporter gene expression (100-1000-fold). Metabolic labeling of the cell medium showed that especially [35S]sulfate-labeled macromolecules competed with DNA for binding to the cationic lipid. Release of DNA from the cationic lipid by cell medium components was demonstrated by an ethidium bromide intercalation assay. In the presence of the cationic lipid, the secreted macromolecules were internalized by the cells. By enzymatic digestions, it was shown that the competing macromolecules consist of chondroitin/dermatan sulfate and heparan sulfate proteoglycans and that the effects on transfection were mediated by the polyanionic glycosaminoglycan portion of the proteoglycan. Accordingly, pretreatment of cell medium with the polycationic peptide protamine sulfate abrogated the inhibitory effects on gene transfer. Fluorescence microscopy studies revealed that heparan sulfate, internalized as a complex with cationic lipids, accumulated in the cell nuclei. These results support the view that the lack of specificity of this type of gene transfer vehicle is a major hindrance to efficient and safe in vivo administration.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
A. Wittrup, S. Sandgren, J. Lilja, C. Bratt, N. Gustavsson, M. Morgelin, and M. Belting
Identification of Proteins Released by Mammalian Cells That Mediate DNA Internalization through Proteoglycan-dependent Macropinocytosis
J. Biol. Chem., September 21, 2007; 282(38): 27897 - 27904.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
L. S. Jones, B. Yazzie, and C. R. Middaugh
Polyanions and the Proteome
Mol. Cell. Proteomics, August 1, 2004; 3(8): 746 - 769.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Sandgren, A. Wittrup, F. Cheng, M. Jonsson, E. Eklund, S. Busch, and M. Belting
The Human Antimicrobial Peptide LL-37 Transfers Extracellular DNA Plasmid to the Nuclear Compartment of Mammalian Cells via Lipid Rafts and Proteoglycan-dependent Endocytosis
J. Biol. Chem., April 23, 2004; 279(17): 17951 - 17956.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
V. P. Torchilin, T. S. Levchenko, R. Rammohan, N. Volodina, B. Papahadjopoulos-Sternberg, and G. G. M. D'Souza
Cell transfection in vitro and in vivo with nontoxic TAT peptide-liposome-DNA complexes
PNAS, February 18, 2003; 100(4): 1972 - 1977.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
F. Cheng, K. Mani, J. van den Born, K. Ding, M. Belting, and L.-A. Fransson
Nitric Oxide-dependent Processing of Heparan Sulfate in Recycling S-Nitrosylated Glypican-1 Takes Place in Caveolin-1-containing Endosomes
J. Biol. Chem., November 8, 2002; 277(46): 44431 - 44439.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Sandgren, F. Cheng, and M. Belting
Nuclear Targeting of Macromolecular Polyanions by an HIV-Tat Derived Peptide. ROLE FOR CELL-SURFACE PROTEOGLYCANS
J. Biol. Chem., October 4, 2002; 277(41): 38877 - 38883.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Durocher, S. Perret, and A. Kamen
High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells
Nucleic Acids Res., January 15, 2002; 30(2): e9 - e9.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. M. Wiethoff, J. G. Smith, G. S. Koe, and C. R. Middaugh
The Potential Role of Proteoglycans in Cationic Lipid-mediated Gene Delivery. STUDIES OF THE INTERACTION OF CATIONIC LIPID-DNA COMPLEXES WITH MODEL GLYCOSAMINOGLYCANS
J. Biol. Chem., August 24, 2001; 276(35): 32806 - 32813.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Ruponen, S. Ronkko, P. Honkakoski, J. Pelkonen, M. Tammi, and A. Urtti
Extracellular Glycosaminoglycans Modify Cellular Trafficking of Lipoplexes and Polyplexes
J. Biol. Chem., August 31, 2001; 276(36): 33875 - 33880.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 1999 by the American Society for Biochemistry and Molecular Biology.