title: Enthalpy-driven interactions with sulfated glycosaminoglycans promote cell membrane penetration of arginine peptides.
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.
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shoumachi, Tokushima 770-8505, Japan; Japan Agency for Medical Research and Development, 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan; Divisioin of Drugs, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Takechi-Haraya
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shoumachi, Tokushima 770-8505, Japan; Department of Biophysical Chemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Nadai
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shoumachi, Tokushima 770-8505, Japan; Department of Biophysical Chemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kimura
authors.affiliation: Department of Human Pathology, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramotocho, Tokushima 770-8503, Japan.
authors.forename: Kazuchika
authors.initials: K
authors.lastname: Nishitsuji
authors.affiliation: Department of Biochemistry, Nagoya University Graduate School of Medicine, 65 Tsurumaicho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Uchimura
authors.affiliation: Divisioin of Drugs, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Sakai-Kato
authors.affiliation: International Center for Materials Nanoarchitectonics, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan.
authors.forename: Kohsaku
authors.initials: K
authors.lastname: Kawakami
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shoumachi, Tokushima 770-8505, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Shigenaga
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Kawakami
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shoumachi, Tokushima 770-8505, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Otaka
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Hironobu
authors.initials: H
authors.lastname: Hojo
authors.affiliation: Department of Human Pathology, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramotocho, Tokushima 770-8503, Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Sakashita
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shoumachi, Tokushima 770-8505, Japan; Department of Biophysical Chemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan. Electronic address: hsaito@mb.kyoto-phu.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Saito
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D016466:CHO Cells; D002463:Cell Membrane Permeability; D006224:Cricetinae; D003412:Cricetulus; D006025:Glycosaminoglycans / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D066244:Proton Magnetic Resonance Spectroscopy; D013431:Sulfates / Q000737:chemistry*; D013816:Thermodynamics; D053835:Unilamellar Liposomes
chemical_list: D006025:Glycosaminoglycans; D010455:Peptides; D013431:Sulfates; D053835:Unilamellar Liposomes; D006493:Heparin; D001120:Arginine
keywords: Arginine peptide; Binding enthalpy; Cell membrane penetration; Glycosaminoglycan; Heparin; Lysine peptide