Soluble Glycosaminoglycans Inhibit the Interaction of TAT−PTD with Lipid Vesicles
- 98 Downloads
Several models have been proposed for translocation of cell-penetrating peptides across membranes, but no general consensus on the mechanism of this process has emerged. It was hypothesized that heparan sulfate on the cell surface may play a role. We used fluorescence spectroscopy to study the effect of three soluble glycosaminoglycans—heparan sulfate, low-molecular-weight heparin, and dermatan sulfate—on the interaction of the fluorescently labeled peptide TAT−PTD with negatively charged small unilamellar vesicles. We found that the presence of glycosaminoglycans results in an order-of-magnitude increase in the apparent dissociation constant K d of the electrostatic component of the peptide/membrane interaction (from 0.13 to 2.6 mM). Thus, rather than aiding in the peptide’s penetration, soluble glycosaminoglycans competitively decrease TAT−PTD’s binding to the membrane, presumably by neutralizing its charge, and thereby attenuating electrostatic forces involved in the interaction. Our results, however, do not exclude a possible role of membrane-anchored glycosaminoglycans in the endocytotic transduction of CPPs across the cell membrane.
KeywordsCell-penetrating peptides Glycosaminoglycans Membrane binding Fluorescence spectroscopy
- Oehlke J, Scheller A, Wiesner B, Krause E, Beyermann M, Klauschenz E, Melzig M, Bienert M (1998) Cellular uptake of an alpha-helical amphipathic model peptide with the potential to deliver polar compounds into the cell interior non-endocytically. Biochim Biophys Acta 1414:127–139PubMedCrossRefGoogle Scholar