title: Anti-heparan sulfate peptides that block herpes simplex virus infection in vivo.
abstract: Heparan sulfate (HS) and its highly modified form, 3-O-sulfated heparan sulfate (3-OS HS), contribute strongly to herpes simplex virus type-1 (HSV-1) infection in vitro. Here we report results from a random M13-phage display library screening to isolate 12-mer peptides that bind specifically to HS, 3-OS HS, and block HSV-1 entry. The screening identified representative candidates from two-different groups of anti-HS peptides with high positive charge densities. Group 1, represented by G1 peptide (LRSRTKIIRIRH), belongs to a class with alternating charges (XRXRXKXXRXRX), and group 2, represented by G2 peptide (MPRRRRIRRRQK), shows repetitive charges (XXRRRRXRRRXK). Viral entry and glycoprotein D binding assays together with fluorescent microscopy data indicated that both G1 and G2 were potent in blocking HSV-1 entry into primary cultures of human corneal fibroblasts and CHO-K1 cells transiently expressing different glycoprotein D receptors. Interestingly, G2 peptide isolated against 3-OS HS displayed wider ability to inhibit entry of clinically relevant strains of HSV-1 and some divergent members of herpesvirus family including cytomegalovirus and human herpesvirus-8. To identify functional residues within G1 and G2, we performed point mutations and alanine-scanning mutagenesis. Several arginine and a lysine residues were needed for anti-HSV-1 activity, suggesting the importance of the positively charged residues in virus-cell binding and virus-induced membrane fusion. In vivo administration of G1 or G2 peptide as a prophylactic eye drop completely blocked HSV-1 spread in the mouse cornea as evident by immunohistochemistry. This result also highlights an in vivo significance of HS and 3-OS HS during ocular herpes infection.
authors: Departments of Ophthalmology and Visual Sciences, University of Illinois, Chicago, Illinois 60612, USA.
authors: Vaibhav
authors: V
authors: Tiwari
authors: Jian
authors: J
authors: Liu
authors: Tibor
authors: T
authors: Valyi-Nagy
authors: Deepak
authors: D
authors: Shukla
references: Virology. 2002 Oct 25;302(2):413-9
references: 12441085
references: J Gen Virol. 2007 Apr;88(Pt 4):1075-1079
references: 17374750
references: Med Res Rev. 2002 Jan;22(1):1-25
references: 11746174
references: J Gen Virol. 2004 Apr;85(Pt 4):805-809
references: 15039523
references: FEBS J. 2009 Dec;276(24):7228-36
references: 19878306
references: Biochem Biophys Res Commun. 2010 Jan 1;391(1):176-81
references: 19909728
references: J Virol. 1991 Mar;65(3):1090-8
references: 1847438
references: Biochem Biophys Res Commun. 2009 Dec 18;390(3):382-7
references: 19747451
references: Biochemistry. 2007 Nov 13;46(45):13049-58
references: 17944486
references: Virology. 2010 Feb 20;397(2):389-98
references: 20004926
references: Trends Microbiol. 2006 Mar;14(3):141-7
references: 16460941
references: FEBS J. 2008 Nov;275(21):5272-85
references: 18803666
references: J Biol Chem. 2007 Sep 21;282(38):27913-22
references: 17640876
references: Cell. 1999 Oct 1;99(1):13-22
references: 10520990
references: J Cell Biol. 1992 Mar;116(5):1273-81
references: 1310996
references: Invest Ophthalmol Vis Sci. 2008 Sep;49(9):4026-35
references: 18502984
references: FEBS Lett. 2007 Sep 18;581(23):4468-72
references: 17765228
references: Immunobiology. 2007;212(9-10):679-91
references: 18086370
references: J Virol. 2001 Feb;75(3):1378-86
references: 11152511
references: J Virol. 2006 Sep;80(18):8970-80
references: 16940509
references: Biochemistry. 2008 May 27;47(21):5774-83
references: 18457417
references: J Virol. 1998 Sep;72(9):7563-8
references: 9696854
references: Virology. 2006 Mar 15;346(2):452-9
references: 16336986
references: J Biol Chem. 2009 Oct 23;284(43):29654-65
references: 19726670
references: J Exp Med. 2009 Nov 23;206(12):2717-33
references: 19858326
references: J Cell Biol. 2006 Sep 25;174(7):1009-21
references: 17000878
references: Virology. 2010 Oct 25;406(2):280-5
references: 20705311
references: Virology. 2001 Jan 5;279(1):313-24
references: 11145912
references: J Virol. 2003 Oct;77(19):10179-85
references: 12970403
references: Virology. 1994 Feb 15;199(1):67-80
references: 8116256
references: Biochem Biophys Res Commun. 2005 Dec 16;338(2):930-7
references: 16259945
references: Nat Chem Biol. 2009 Oct;5(10):743-8
references: 19734912
references: J Virol. 1989 Jan;63(1):52-8
references: 2535752
references: J Clin Invest. 2001 Aug;108(4):503-10
references: 11518721
references: J Gen Virol. 1991 May;72 ( Pt 5):1131-7
references: 1851813
references: Nature. 2007 Apr 26;446(7139):1030-7
references: 17460664
references: Cell. 1996 Nov 1;87(3):427-36
references: 8898196
references: Biochem J. 2005 Jan 15;385(Pt 2):451-9
references: 15303968
references: Antimicrob Agents Chemother. 2010 Oct;54(10):4290-9
references: 20643894
references: Cornea. 2001 Jan;20(1):1-13
references: 11188989
references: J Biol Chem. 2002 Oct 4;277(40):37912-9
references: 12138164
references: J Clin Invest. 2001 Jul;108(2):169-73
references: 11457867
references: Antiviral Res. 2010 Mar;85(3):532-40
references: 20083141