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.
Department of Clinical Sciences, Section of Oncology, Lund University, Lund, Sweden.
Anders
A
Wittrup
Si-He
SH
Zhang
Mattias
M
Belting
D000818:Animals; D016466:CHO Cells; D057846:Cell-Penetrating Peptides / Q000302:isolation & purification / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D004983:Ethanolamines / Q000737:chemistry; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D021381:Protein Transport; D011509:Proteoglycans / Q000378:metabolism*; D053669:Syndecan-2 / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics / Q000302:isolation & purification
D057846:Cell-Penetrating Peptides; D004983:Ethanolamines; D011509:Proteoglycans; D053669:Syndecan-2; C007369:2-diethylaminoethanol