Title
Enthalpy-driven interactions with sulfated glycosaminoglycans promote cell membrane penetration of arginine peptides
Author
Yuki Takechi-Haraya, Ryo Nadai, Hitoshi Kimura, Kazuchika Nishitsuji, Kenji Uchimura, Kumiko Sakai-Kato, Kohsaku Kawakami, Akira Shigenaga, Toru Kawakami, Akira Otaka, Hironobu Hojo, Naomi Sakashita, Hiroyuki Saito
Year
2016
Journal
Biochimica et Biophysica Acta-Biomembranes
Abstract
The first step of cell membrane penetration of arginine peptides is thought to occur via electrostatic interactions between positive charges of arginine residues and negative charges of sulfated glycosaminoglycans (GAGs) on the cell surface. However, the molecular interaction of arginine peptides with GAG still remains unclear. Here, we compared the interactions of several arginine peptides of Tat, R8, and Rev and their analogues with heparin in relation to the cell membrane penetration efficiency. The high-affinity binding of arginine peptides to heparin was shown to be driven by large favorable enthalpy contributions, possibly reflecting multidentate hydrogen bondings of arginine residues with sulfate groups of heparin. Interestingly, the lysine peptides in which all arginine residues are substituted with lysine residues exhibited negligible binding enthalpy despite of their considerable binding to heparin. In CHO-K1 cells, arginine peptides exhibited a great cell-penetrating ability whereas their corresponding lysine peptides did not penetrate into cells. The degree of cell penetration of arginine peptides markedly decreased by the chlorate treatment of cells which prevents the sulfation of GAG chains. Significantly, the cell penetration efficiency of arginine peptides was found to be correlated with the favorable enthalpy of binding to heparin. These results suggest that the enthalpy-driven strong interaction with sulfated GAGs such as heparan sulfate plays a critical role in the efficient cell membrane penetration of arginine peptides.
Instrument
J-1500
Keywords
Circular dichroism, Secondary structure, Ligand binding, Biochemistry