Heparan sulfate proteoglycan-mediated polyamine uptake.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1831860
- author
- Welch, Johanna LU ; Svensson, Katrin LU ; Kucharzewska, Paulina LU and Belting, Mattias LU
- organization
- publishing date
- 2011
- 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
- pmid:21318883
- 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
- 2016-04-04 07:27:19
- date last changed
- 2022-01-29 02:11:18
@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}}, doi = {{10.1007/978-1-61779-034-8_20}}, volume = {{720}}, year = {{2011}}, }