title: Selective Intracellular Delivery of Ganglioside GM3-Binding Peptide through Caveolae/Raft-Mediated Endocytosis.
abstract: Glycosphingolipids are major components of the membrane raft, and several kinds of viruses and bacterial toxins are known to bind to glycosphingolipids in the membrane raft. Since the viral genes and pathogenic proteins that are taken into cells are directly delivered to their target organelles, caveolae/raft-mediated endocytosis represents a promising pathway for specific delivery. In the present study, we demonstrated the ability of an artificial pentadecapeptide, which binds to ganglioside GM3, to deliver protein into cells by caveolae/raft-mediated endocytosis. The cellular uptake of a biotinylated GM3-binding peptide (GM3BP)-avidin complex into HeLa cells was observed, and the cellular uptake of this complex was inhibited by an incubation with sialic acid or endocytic inhibitors such as methyl-ß-cyclodextrin, and also by an incubation at 4 °C. These results indicate that the GM3BP-avidin complex bind to GM3 in membrane raft, and are taken into cell through caveolae/raft-mediated endocytosis. The GM3BP-avidin complex was transported into cells and localized around the nucleus more slowly than a human immunodeficiency virus type 1 TAT peptide. Furthermore, the uptake of a green fluorescent protein (GFP) linked with GM3BP into HeLa cells was similar to that of the GM3BP-avidin complex, and the localization of the GM3BP-GFP fusion protein was markedly different with that of the TAT-GFP fusion protein. The uptake and trafficking of GM3BP were distinguished from conventional cell-penetrating peptides. GM3BP has potential as a novel peptide for the selective delivery of therapeutic proteins and materials into cells in addition to being a cell-penetrating peptide.
authors.affiliation: Department of Biosciences and Informatics, Keio University , 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama 223-8522, Japan.
authors.forename: Teruhiko
authors.identifier: 0000-0002-8006-4324
authors.initials: T
authors.lastname: Matsubara
authors.affiliation: Department of Biosciences and Informatics, Keio University , 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama 223-8522, Japan.
authors.forename: Ryohei
authors.initials: R
authors.lastname: Otani
authors.affiliation: Department of Biosciences and Informatics, Keio University , 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama 223-8522, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Yamashita
authors.affiliation: Department of Biosciences and Informatics, Keio University , 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama 223-8522, Japan.
authors.forename: Haruka
authors.initials: H
authors.lastname: Maeno
authors.affiliation: Department of Biosciences and Informatics, Keio University , 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama 223-8522, Japan.
authors.forename: Hanae
authors.initials: H
authors.lastname: Nodono
authors.affiliation: Department of Biosciences and Informatics, Keio University , 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama 223-8522, Japan.
authors.forename: Toshinori
authors.initials: T
authors.lastname: Sato
mesh_terms: D000818:Animals; D019556:COS Cells; D021941:Caveolae / Q000378:metabolism*; D002470:Cell Survival; D057846:Cell-Penetrating Peptides / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D016503:Drug Delivery Systems; D004705:Endocytosis / Q000502:physiology*; D005679:G(M3) Ganglioside / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space; D021962:Membrane Microdomains / Q000378:metabolism*; D021381:Protein Transport; D015398:Signal Transduction; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D057846:Cell-Penetrating Peptides; D005679:G(M3) Ganglioside; D054322:tat Gene Products, Human Immunodeficiency Virus