title: Cell-surface accumulation of flock house virus-derived peptide leads to efficient internalization via macropinocytosis.
abstract: Arginine-rich cell-penetrating peptides (CPPs), including human immunodeficiency virus type 1 (HIV-1) Tat (48-60) and oligoarginines, have been applied as carriers for delivery of cargo molecules, because of their capacity to internalize into cells and penetrate biological membranes. Despite the fact that they have been extensively studied, the factors required for the efficient internalization of CPPs are still unclear. In this report, we evaluated the internalization efficiencies of seven CPPs derived from DNA/RNA-binding peptides, and discovered that a peptide derived from the flock house virus (FHV) coat protein was internalized most efficiently into Chinese hamster ovary (CHO-K1), HeLa, and Jurkat cells. Comparison of the factors facilitating the internalization with those of the Tat peptide revealed that the FHV peptide induces macropinocytosis much more efficiently than the Tat peptide, which leads to its high cellular uptake efficiency. Additionally, the strong adsorption of the FHV peptide on cell membranes via glycosaminoglycans (GAGs) was shown to be a key factor for induction of macropinocytosis, and these steps were successfully monitored by live imaging of the peptide internalization into cells in relation to the actin organization. The remarkable methods of FHV peptide internalization thus highlighted the critical factors for internalizations of the arginine-rich CPPs.
authors.affiliation: Institute for Chemical Research, Kyoto University, Kyoto, Japan.
authors.forename: Ikuhiko
authors.initials: I
authors.lastname: Nakase
authors.forename: Hisaaki
authors.initials: H
authors.lastname: Hirose
authors.forename: Gen
authors.initials: G
authors.lastname: Tanaka
authors.forename: Akiko
authors.initials: A
authors.lastname: Tadokoro
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kobayashi
authors.forename: Toshihide
authors.initials: T
authors.lastname: Takeuchi
authors.forename: Shiroh
authors.initials: S
authors.lastname: Futaki
mesh_terms: D000818:Animals; D016466:CHO Cells; D036022:Capsid Proteins / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D015638:Cytochalasin D / Q000494:pharmacology; D005434:Flow Cytometry; D006025:Glycosaminoglycans / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010446:Peptide Fragments / Q000378:metabolism*; D010873:Pinocytosis / Q000187:drug effects / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D006025:Glycosaminoglycans; D019384:Nucleic Acid Synthesis Inhibitors; D010446:Peptide Fragments; D054322:tat Gene Products, Human Immunodeficiency Virus; D015638:Cytochalasin D
references.citation: Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13683-8
references.pmid: 9391086
references.citation: J Cell Mol Med. 2007 Jul-Aug;11(4):670-84
references.pmid: 17760832
references.citation: J Cell Biol. 1998 Jul 13;142(1):139-51
references.pmid: 9660869
references.citation: J Biol Chem. 2001 Feb 23;276(8):5836-40
references.pmid: 11084031
references.citation: J Cell Biol. 1998 Jul 13;142(1):129-38
references.pmid: 9660868
references.citation: J Cell Sci. 1992 Aug;102 ( Pt 4):867-80
references.pmid: 1429898
references.citation: J Biol Chem. 1997 Jun 20;272(25):16010-7
references.pmid: 9188504
references.citation: J Biol Chem. 2003 Jan 3;278(1):585-90
references.pmid: 12411431
references.citation: Mol Ther. 2004 Dec;10(6):1011-22
references.pmid: 15564133
references.citation: J Cell Biol. 1996 Dec;135(5):1249-60
references.pmid: 8947549
references.citation: Trends Cell Biol. 1995 Nov;5(11):424-8
references.pmid: 14732047
references.citation: Exp Cell Res. 1998 Aug 1;242(2):528-39
references.pmid: 9683539
references.citation: Biochemistry. 2004 Mar 9;43(9):2438-44
references.pmid: 14992581
references.citation: J Cell Sci. 2006 Jul 1;119(Pt 13):2837-46
references.pmid: 16787950
references.citation: Curr Opin Cell Biol. 2001 Oct;13(5):578-83
references.pmid: 11544026
references.citation: Microbiol Immunol. 2004;48(11):823-9
references.pmid: 15557740
references.citation: Cell. 1993 Apr 23;73(2):309-19
references.pmid: 8477447
references.citation: Circ Res. 2006 Jun 9;98(11):1398-404
references.pmid: 16675718
references.citation: Nat Med. 2004 Mar;10(3):310-5
references.pmid: 14770178
references.citation: Biol Cell. 2007 Apr;99(4):223-38
references.pmid: 17233629
references.citation: Traffic. 2007 Jul;8(7):848-66
references.pmid: 17587406
references.citation: Biochem J. 2006 Aug 15;398(1):1-13
references.pmid: 16856876
references.citation: J Cell Biol. 2002 Sep 16;158(6):1119-31
references.pmid: 12221069
references.citation: J Biol Chem. 2005 Apr 15;280(15):15300-6
references.pmid: 15687490
references.citation: J Biol Chem. 2001 Feb 2;276(5):3254-61
references.pmid: 11024024
references.citation: Science. 2008 Apr 25;320(5875):531-5
references.pmid: 18436786
references.citation: J Virol. 2001 Nov;75(22):11166-77
references.pmid: 11602756
references.citation: J Control Release. 2005 Jan 20;102(1):247-53
references.pmid: 15653149
references.citation: Matrix Biol. 2003 Mar;22(1):25-33
references.pmid: 12714039
references.citation: J Biol Chem. 2002 Jan 25;277(4):2437-43
references.pmid: 11711547
references.citation: Infect Immun. 1996 May;64(5):1621-30
references.pmid: 8613370
references.citation: J Cell Sci. 2006 Nov 15;119(Pt 22):4758-69
references.pmid: 17077125
references.citation: Nat Rev Mol Cell Biol. 2008 Aug;9(8):639-49
references.pmid: 18612320
references.citation: Bioconjug Chem. 2008 Mar;19(3):656-64
references.pmid: 18269225
references.citation: Cell Microbiol. 2000 Feb;2(1):69-82
references.pmid: 11207564
references.citation: Nat Med. 2000 Nov;6(11):1253-7
references.pmid: 11062537
references.citation: Biochemistry. 1991 Feb 19;30(7):1973-80
references.pmid: 1899622
references.citation: Biopolymers. 2007;88(2):98-107
references.pmid: 17252560
references.citation: Biopolymers. 2006;84(3):241-9
references.pmid: 16333858
references.citation: Biochem J. 2007 Apr 15;403(2):335-42
references.pmid: 17217340
references.citation: Curr Med Chem. 2008;15(19):1940-55
references.pmid: 18691050
references.citation: J Virol. 2006 Dec;80(23):11833-51
references.pmid: 16987972
references.citation: J Cell Biol. 2007 May 7;177(3):527-38
references.pmid: 17485492
references.citation: Biophys J. 2004 Jan;86(1 Pt 1):254-63
references.pmid: 14695267
references.citation: Biochemistry. 2007 Jan 16;46(2):492-501
references.pmid: 17209559
references.citation: Bioconjug Chem. 2004 May-Jun;15(3):475-81
references.pmid: 15149174
references.citation: Biophys J. 2008 Mar 15;94(6):2142-9
references.pmid: 18065465
references.citation: Biochem Biophys Res Commun. 2007 Nov 30;363(4):1027-32
references.pmid: 17923117
references.citation: Adv Drug Deliv Rev. 2008 Mar 1;60(4-5):447
references.pmid: 18077052