title: Cell-penetrating compounds preferentially bind glycosaminoglycans over plasma membrane lipids in a charge density- and stereochemistry-dependent manner.
abstract: Cell-penetrating compounds (CPCs) are often conjugated to drugs and genes to facilitate cellular uptake. We hypothesize that the electrostatic interaction between the positively charged amines of the cell-penetrating compounds and the negatively charged glycosaminoglycans (GAGs) extending from cell surfaces is the initiating step in the internalization process. The interactions of generation 5 PAMAM dendrimer, Tat peptide and 25 kDa linear PEI with four different GAGs have been studied using isothermal titration calorimetry to elucidate structure-function relationships that could lead to improved drug and gene delivery methods to a wide variety of cell types. Detailed thermodynamic analysis has determined that CPC-GAG binding constants range from 8.7×10(3) to 2.4×10(6)M(-1) and that affinity is dependent upon GAG charge density and stereochemistry and CPC molecular weight. The effect of GAG composition on affinity is likely due to hydrogen bonding between CPC amines and amides and GAG hydroxyl and amine groups. These results were compared to the association of CPCs with lipid vesicles of varying composition as model plasma membranes to finally clarify the relative importance of each cell surface component in initial cell recognition. CPC-lipid affinity increases with anionic lipid content, but GAG affinity is higher for all cell-penetrating compounds, confirming the role these heterogeneous polysaccharides play in cellular association and clustering.
authors.affiliation: Department of Chemistry, University of St. Thomas, 2115 Summit Ave., St. Paul, MN 55105-1079, United States. Electronic address: lisa.prevette@stthomas.edu.
authors.forename: Lisa E
authors.initials: LE
authors.lastname: Prevette
authors.affiliation: Department of Chemistry, University of St. Thomas, 2115 Summit Ave., St. Paul, MN 55105-1079, United States.
authors.forename: Nicolas C
authors.initials: NC
authors.lastname: Benish
authors.affiliation: Department of Chemistry, University of St. Thomas, 2115 Summit Ave., St. Paul, MN 55105-1079, United States.
authors.forename: Amber R
authors.initials: AR
authors.lastname: Schoenecker
authors.affiliation: Department of Chemistry, University of St. Thomas, 2115 Summit Ave., St. Paul, MN 55105-1079, United States.
authors.forename: Kristin J
authors.initials: KJ
authors.lastname: Braden
mesh_terms: D002151:Calorimetry; D050091:Dendrimers / Q000737:chemistry; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D050091:Dendrimers; D006025:Glycosaminoglycans; D008051:Lipid Bilayers; D008081:Liposomes; C104700:PAMAM Starburst
keywords: Cell surface interactions; Cell-penetrating compounds; Cell-penetrating peptides; Electrostatic free energy; Glycosaminoglycans; Isothermal titration calorimetry; Lipid vesicles