title: Modular analysis of hipposin, a histone-derived antimicrobial peptide consisting of membrane translocating and membrane permeabilizing fragments.
abstract: Antimicrobial peptides continue to garner attention as potential alternatives to conventional antibiotics. Hipposin is a histone-derived antimicrobial peptide (HDAP) previously isolated from Atlantic halibut. Though potent against bacteria, its antibacterial mechanism had not been characterized. The mechanism of this peptide is particularly interesting to consider since the full hipposin sequence contains the sequences of parasin and buforin II (BF2), two other known antimicrobial peptides that act via different antibacterial mechanisms. While parasin kills bacteria by inducing membrane permeabilization, buforin II enters cells without causing significant membrane disruption, harming bacteria through interactions with intracellular nucleic acids. In this study, we used a modular approach to characterize hipposin and determine the role of the parasin and buforin II fragments in the overall hipposin mechanism. Our results show that hipposin kills bacteria by inducing membrane permeabilization, and this membrane permeabilization is promoted by the presence of the N-terminal domain. Portions of hipposin lacking the N-terminal sequence do not cause membrane permeabilization and function more similarly to buforin II. We also determined that the C-terminal portion of hipposin, HipC, is a cell-penetrating peptide that readily enters bacterial cells but has no measurable antimicrobial activity. HipC is the first membrane active histone fragment identified that does not kill bacterial or eukaryotic cells. Together, these results characterize hipposin and provide a useful starting point for considering the activity of chimeric peptides made by combining peptides with different antimicrobial mechanisms. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova.
authors.affiliation: Department of Chemistry, Wellesley College, Wellesley, MA 02481.|Department of Biological Sciences, Wellesley College, Wellesley, MA 02481.
authors.forename: Maria E
authors.initials: ME
authors.lastname: Bustillo
authors.affiliation: Department of Chemistry, Wellesley College, Wellesley, MA 02481.
authors.forename: Alexandra L
authors.initials: AL
authors.lastname: Fischer
authors.affiliation: Department of Chemistry, Wellesley College, Wellesley, MA 02481.|Department of Biological Sciences, Wellesley College, Wellesley, MA 02481.
authors.forename: Maria A
authors.initials: MA
authors.lastname: LaBouyer
authors.affiliation: Department of Chemistry, Wellesley College, Wellesley, MA 02481.
authors.forename: Julia A
authors.initials: JA
authors.lastname: Klaips
authors.affiliation: Department of Biological Sciences, Wellesley College, Wellesley, MA 02481.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Webb
authors.affiliation: Department of Chemistry, Wellesley College, Wellesley, MA 02481.
authors.forename: Donald E
authors.initials: DE
authors.lastname: Elmore
mesh_terms: D000890:Anti-Infective Agents / Q000737:chemistry* / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000494:pharmacology; D002942:Circular Dichroism; D029941:Fish Proteins / Q000737:chemistry* / Q000494:pharmacology; D006657:Histones / Q000737:chemistry; D008826:Microbial Sensitivity Tests; D011506:Proteins / Q000737:chemistry* / Q000494:pharmacology
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D057846:Cell-Penetrating Peptides; D029941:Fish Proteins; D006657:Histones; D011506:Proteins; C097846:buforin I; C097849:buforin II; C476394:hipposin protein, Hippoglossus hippoglossus; C115701:parasin I
keywords: Buforin; Hipposin; Histone-derived antimicrobial peptide; Membrane permeabilization; Membrane translocation; Parasin
references.citation: Methods Mol Biol. 2011;683:21-9
references.pmid: 21053119
references.citation: PLoS Pathog. 2010 Apr 22;6(4):e1000857
references.pmid: 20421939
references.citation: Infect Disord Drug Targets. 2008 Sep;8(3):195-205
references.pmid: 18782037
references.citation: Biochim Biophys Acta. 2009 Aug;1788(8):1564-9
references.pmid: 19041293
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D933-7
references.pmid: 18957441
references.citation: Biochim Biophys Acta. 2012 Mar;1818(3):869-76
references.pmid: 22230351
references.citation: J Immunol Methods. 1991 Mar 21;137(2):167-73
references.pmid: 1901580
references.citation: J Control Release. 2012 Nov 10;163(3):293-303
references.pmid: 23041543
references.citation: Curr Drug Targets Infect Disord. 2002 Mar;2(1):79-83
references.pmid: 12462155
references.citation: Proteins. 2008 Nov 1;73(2):480-91
references.pmid: 18452210
references.citation: Trends Biotechnol. 2011 Sep;29(9):464-72
references.pmid: 21680034
references.citation: FEBS Lett. 1998 Oct 23;437(3):258-62
references.pmid: 9824303
references.citation: J Vis Exp. 2012 Jan 27;(59):e3571
references.pmid: 22314806
references.citation: J Control Release. 2012 Jul 20;161(2):582-91
references.pmid: 22516088
references.citation: Biochim Biophys Acta. 2003 Mar 21;1646(1-2):207-15
references.pmid: 12637028
references.citation: Biochemistry. 2010 Sep 14;49(36):7854-66
references.pmid: 20704453
references.citation: Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8245-50
references.pmid: 10890923
references.citation: Biochim Biophys Acta. 1998 Nov 10;1376(3):391-400
references.pmid: 9804997
references.citation: Peptides. 2011 Apr;32(4):677-82
references.pmid: 21277926
references.citation: Biochem Biophys Res Commun. 1996 Jan 5;218(1):408-13
references.pmid: 8573171
references.citation: Biochemistry. 2004 Dec 14;43(49):15610-6
references.pmid: 15581374
references.citation: Antimicrob Agents Chemother. 2011 Mar;55(3):954-60
references.pmid: 21189349
references.citation: Biochemistry. 2000 Jul 25;39(29):8648-54
references.pmid: 10913273
references.citation: Biochim Biophys Acta. 2012 Mar;1818(3):762-75
references.pmid: 22155682
references.citation: Biochemistry. 2001 Dec 4;40(48):14330-5
references.pmid: 11724544
references.citation: Biochem Biophys Res Commun. 1998 Mar 6;244(1):253-7
references.pmid: 9514864
references.citation: FEBS Lett. 2008 May 28;582(12):1715-8
references.pmid: 18448075
references.citation: Biochim Biophys Acta. 2004 Jun 1;1699(1-2):221-7
references.pmid: 15158731
references.citation: Peptides. 2008 Jul;29(7):1102-8
references.pmid: 18406495
references.citation: Peptides. 2009 May;30(5):839-48
references.pmid: 19428759
references.citation: Mar Drugs. 2010 Apr 14;8(4):1213-62
references.pmid: 20479976
references.citation: Peptides. 2009 Dec;30(12):2168-73
references.pmid: 19770014
references.citation: J Biol Chem. 2001 Dec 14;276(50):47070-7
references.pmid: 11598107