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Heparan sulfate proteoglycan-mediated polyamine uptake.

Welch, Johanna LU ; Svensson, Katrin LU ; Kucharzewska, Paulina LU and Belting, Mattias LU (2011) In Methods in Molecular Biology 720. p.327-338
Abstract
The polyamines are polycationic compounds essential for cellular proliferation and transformation. In addition to a well-defined biosynthesis pathway, polyamines are internalized into cells by as yet incompletely defined mechanisms. Numerous reports have shown that efficient polyamine uptake depends on the presence of polyanionic, cell surface-associated heparan sulfate proteoglycans (HSPGs). In this chapter, we provide protocols for studying HSPG-mediated uptake of polyamines in various cell lines, and provide instructions for the use of two different genetic models of HSPG deficiency. We describe the enzymatic reduction of cell surface HSPG through Heparinase III lyase treatment as well as the use of phage display-derived single chain... (More)
The polyamines are polycationic compounds essential for cellular proliferation and transformation. In addition to a well-defined biosynthesis pathway, polyamines are internalized into cells by as yet incompletely defined mechanisms. Numerous reports have shown that efficient polyamine uptake depends on the presence of polyanionic, cell surface-associated heparan sulfate proteoglycans (HSPGs). In this chapter, we provide protocols for studying HSPG-mediated uptake of polyamines in various cell lines, and provide instructions for the use of two different genetic models of HSPG deficiency. We describe the enzymatic reduction of cell surface HSPG through Heparinase III lyase treatment as well as the use of phage display-derived single chain variable fragment (scFv) anti-HS antibodies to block HSPGs at the cell surface. Finally, we provide a protocol for the quantitative verification of loss or reduction of cell surface HSPGs and a detailed description of polyamine uptake measurement. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Methods in Molecular Biology
volume
720
pages
327 - 338
publisher
Springer
external identifiers
  • pmid:21318883
  • scopus:79956209167
ISSN
1940-6029
DOI
10.1007/978-1-61779-034-8_20
language
English
LU publication?
yes
id
742173f3-a862-470f-a739-6aa1fd9e28ff (old id 1831860)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/21318883?dopt=Abstract
date added to LUP
2011-03-01 14:53:57
date last changed
2017-01-01 07:31:39
@article{742173f3-a862-470f-a739-6aa1fd9e28ff,
  abstract     = {The polyamines are polycationic compounds essential for cellular proliferation and transformation. In addition to a well-defined biosynthesis pathway, polyamines are internalized into cells by as yet incompletely defined mechanisms. Numerous reports have shown that efficient polyamine uptake depends on the presence of polyanionic, cell surface-associated heparan sulfate proteoglycans (HSPGs). In this chapter, we provide protocols for studying HSPG-mediated uptake of polyamines in various cell lines, and provide instructions for the use of two different genetic models of HSPG deficiency. We describe the enzymatic reduction of cell surface HSPG through Heparinase III lyase treatment as well as the use of phage display-derived single chain variable fragment (scFv) anti-HS antibodies to block HSPGs at the cell surface. Finally, we provide a protocol for the quantitative verification of loss or reduction of cell surface HSPGs and a detailed description of polyamine uptake measurement.},
  author       = {Welch, Johanna and Svensson, Katrin and Kucharzewska, Paulina and Belting, Mattias},
  issn         = {1940-6029},
  language     = {eng},
  pages        = {327--338},
  publisher    = {Springer},
  series       = {Methods in Molecular Biology},
  title        = {Heparan sulfate proteoglycan-mediated polyamine uptake.},
  url          = {http://dx.doi.org/10.1007/978-1-61779-034-8_20},
  volume       = {720},
  year         = {2011},
}