Studies of Proteoglycan Involvement in CPP-Mediated Delivery
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.
Key wordsCell-penetrating peptides CHO cell mutants Heparan sulfate Glypican Proteoglycan siRNA Syndecan
This book chapter is based on work supported by grants from the Swedish Cancer Fund; the Swedish Research Council; the Swedish Foundation for Strategic Research; the Swedish Society of Medicine; the Physiographic Society, Lund; the Crafoordska, Gunnar Nilsson, Lundbergs, and Kamprad Foundations; the Lund University Hospital donation funds; and the governmental funding of clinical research within the National Health Services.
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