AI Article Synopsis
- Cell-penetrating peptides (CPPs) are important tools for delivering large molecules inside cells, and their effectiveness often depends on their interaction with cell surface heparan sulfate proteoglycans (HSPGs).
- The chapter outlines various methods to investigate the role of proteoglycans in the uptake of CPPs, including techniques to assess whether a specific CPP's uptake relies on PGs.
- It also details protocols for purifying PGs and analyzing HSPG protein composition in cell lines, as well as strategies for altering the expression of certain HSPG proteins to study their specificity in CPP uptake.
Article Abstract
Cell-penetrating peptides (CPPs) are widely used to deliver macromolecular cargoes to intracellular sites of action. Many CPPs have been demonstrated to rely on cell surface heparan sulfate proteoglycans (HSPGs) for efficient cellular entry and delivery. In this chapter, we describe methods for the study of PG involvement in CPP uptake. We provide descriptions of how to determine whether uptake of a CPP of interest is dependent on PGs. We also provide detailed protocols for the purification of PGs by anion-exchange chromatography as well as the characterization of the HSPG core protein composition of a cell line of interest. Finally, we present methods for modulating the expression level of specific HSPG core proteins as a means to determine the core protein specificity in the uptake of a particular CPP.
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| http://dx.doi.org/10.1007/978-1-60761-919-2_8 | DOI Listing |
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