Bilayer lipid composition modulates the activity of dermaseptins, polycationic antimicrobial peptides



The primary targets of defense peptides are plasma membranes, and the induced irreversible depolarization is sufficient to exert antimicrobial activity although secondary modes of action might be at work. Channels or pores underlying membrane permeabilization are usually quite large with single-channel conductances two orders of magnitude higher than those exhibited by physiological channels involved, e.g., in excitability. Accordingly, the ion specificity and selectivity are quite low. Whereas, e.g., peptaibols favor cation transport, polycationic or basic peptides tend to form anion-specific pores. With dermaseptin B2, a 33 residue long and mostly α-helical peptide isolated from the skin of the South American frog Phyllomedusa bicolor, we found that the ion specificity of its pores induced in bilayers is modulated by phospholipid-charged headgroups. This suggests mixed lipid–peptide pore lining instead of the more classical barrel–stave model. Macroscopic conductance is nearly voltage independent, and concentration dependence suggests that the pores are mainly formed by dermaseptin tetramers. The two most probable single-channel events are well resolved at 200 and 500 pS (in 150 mM NaCl) with occasional other equally spaced higher or lower levels. In contrast to previous molecular dynamics previsions, this study demonstrates that dermaseptins are able to form pores, although a related analog (B6) failed to induce any significant conductance. Finally, the model of the pore we present accounts for phospholipid headgroups intercalated between peptide helices lining the pore and for one of the most probable single-channel conductance.


Pore-forming peptides Antibiotic resistance Planar lipid bilayers Phospholipids Conductance properties 


  1. Amar B, Perianin A, Mor A, Sarfati G, Tissot M, Nicolas P, Giroud JP, Roch-Arweiller M (1998) Dermaseptin, a peptide antibiotic, stimulates microbicidal activities of poymorphonuclear leukocytes. Biochem Biophys Res Commun 247:870–875CrossRefGoogle Scholar
  2. Balayssac S, Burlina F, Convert O, Bolbach G, Chassaing G, Lequin O (2006) Comparison of penetratin and other homeodomain-derived cell-penetrating peptides: interaction in a membrane-mimicking environment and cellular uptake efficiency. Biochemistry 45(5):1408–1420CrossRefGoogle Scholar
  3. Bechinger B, Aisenbrey C, Bertani P (2005) Detergent-like properties of magainin antibiotic peptides: a 31P solid-state NMR spectroscopy study. Biochim Biophys Acta 1712:101–108CrossRefGoogle Scholar
  4. Béven L, Helluin O, Duclohier H, Wroblewski H (1999) Correlation between antibacterial activity and pore sizes of two classes of voltage-dependent channel-forming peptides. Biochim Biophys Acta 1421:53–63CrossRefGoogle Scholar
  5. Boman HG (2003) Antibacterial peptides: basic facts and emerging concepts. J Intern Med 254:197–215CrossRefGoogle Scholar
  6. Bulet P, Stocklin R (2005) Insect antimicrobial peptides: structures, properties and gene regulation. Protein Pept Lett 12:3–11CrossRefGoogle Scholar
  7. Bulet P, Stocklin R, Menin L (2004) Antimicrobial peptides: from invertebrates to vertebrates. Immunol Rev 198:169–184CrossRefGoogle Scholar
  8. Castiglione-Morelli MA, Cristinziano P, Pepe A, Temussi PA (2005) Conformation–activity relationship of a novel peptide antibiotic: structural characterization of dermaseptin DS 01 in media that mimic the membrane environment. Biopolymers 47:688–696CrossRefGoogle Scholar
  9. Chugh JK, Wallace BA (2001) Peptaibols: models for ion channels. Biochem Soc Trans 29:565–570CrossRefGoogle Scholar
  10. Cruz-Chamorro L, Puertollano MA, Puertollano E, de Cienfuegos GA, de Pablo MA (2005) In vitro biological activities of magainin alone or in combination with nisin. Peptides (in press)Google Scholar
  11. Daly JW, Caceres J, Moni RW, Gusovsky F, Moos M Jr, Seamon KB, Milton K, Myers CW (1992) Frog secretions and hunting magic in the upper Amazon: identification of a peptide that interacts with an adenosine receptor. Proc Natl Acad Sci USA 89:10960–10963CrossRefADSGoogle Scholar
  12. Duclohier H (2002) How do channel- and pore-forming helical peptides interact with lipid membranes and does this account for their antimicrobial activity? Mini Rev Med Chem 2:331–342CrossRefGoogle Scholar
  13. Duclohier H (2003) Insights into ion channels from peptides in planar lipid bilayers. In: Tien HT, Ottova-Leitmannova A (eds) Planar lipid bilayers (BLMs) and their applications. Elsevier, Amsterdam, pp 89–604Google Scholar
  14. Duclohier H, Spach G (2001) Artificial membrane excitability revisited and implications for the gating of voltage-dependent ion channels. Gen Physiol Biophys 20:361–374Google Scholar
  15. Duclohier H, Wroblewski H (2001) Voltage-dependent pore formation and antimicrobial activity by alamethicin and analogues. J Membr Biol 184:1–12CrossRefGoogle Scholar
  16. Duclohier H, Molle G, Spach G (1989) The antimicrobial peptide magainin I from Xenopus skin forms anion-permeable channels in planar lipid bilayers. Biophys J 56:175–188CrossRefGoogle Scholar
  17. Eisenberg M, Hall JE, Mead CA (1973) The nature of the voltage-dependent conductance induced by alamethicin in black lipid membranes. J Membr Biol 14:143–176CrossRefGoogle Scholar
  18. Fleury Y, Vouille V, Beven L, Amiche M, Wroblewski H, Delfour A, Nicolas P (1998) Synthesis, antimicrobial activity and gene structure of a novel member of the dermaseptin B family. Biochim Biophys Acta 1396:228–236Google Scholar
  19. Fox RO, Richards FM (1982) A voltage-gated ion channel inferred from the crystal structure of alamethicin at 1.5 A resolution. Nature 25:325–330CrossRefADSGoogle Scholar
  20. Goldman DE (1943) Potential, impedance, and rectification in membranes. J Gen Physiol 27:37–60CrossRefGoogle Scholar
  21. Hall JE, Vodyanoy I, Balasubramanian TM, Marshall GR (1984) Alamethicin: a rich model for channel behavior. Biophys J 45:233–247Google Scholar
  22. Hanke W, Boheim G (1980) The lowest conductance state of the alamethicin pore. Biochim Biophys Acta 596:456–462CrossRefGoogle Scholar
  23. Hanke W, Methfessel C, Wilmsen U, Boheim G (1984) Ion channel reconstitution into planar lipid bilayers on glass pipettes. Biochem Bioenerg J 12:329–339Google Scholar
  24. Hille B (1992) Ionic channels of excitable membranes, 2nd edn. Chapter 11: Elementary properties of pores. Sinauer Associates, Sunderland, pp 291–314Google Scholar
  25. Hodgkin AL, Katz B (1949) The effect of sodium ions on the electrical activity of the giant axon of the squid. J Physiol 108:37–77Google Scholar
  26. Homblé F, Cabiaux V, Ruysschaert J-M (1998) Channel or channel-like activity associated with pore-forming proteins or peptides. Mol Microbiol 27:1261–1263CrossRefGoogle Scholar
  27. Jones LR, Maddock SW, Besch HR Jr (1980) Unmasking effect of alamethicin on the (Na+-K+)-ATPase, beta-adrenergic receptor-coupled adenylate cyclase, and cAMP-dependent protein kinase activities of cardiac sarcolemmal vesicles. J Biol Chem 255:9971–9980Google Scholar
  28. Keller SL, Bezrukov SM, Gruner SM, Tate MW, Vodyanoy I, Parsegian VA (1993) Probability of alamethicin conductance states varies with nonlamellar tendency of bilayer phospholipids. Biophys J 65:23–27Google Scholar
  29. Kobayashi S, Chikushi A, Tougu S, Imura Y, Yano Y, Matsuzaki K (2004) Membrane translocation mechanism of the antimicrobial peptide buforin 2. Biochem 43:15610–15616CrossRefGoogle Scholar
  30. Kragol G, Lovas S, Varadi G, Condie BA, Hoffmann R (2001) The antibacterial peptide pyrrhocoricin inhibits the ATPase actions of DnaK and prevents chaperone-assisted protein folding. Biochemistry 40:3016–3026CrossRefGoogle Scholar
  31. Krasilnikov OV, Merzlyak PG, Yuldasheva LN, Capistrano MF (2005) Protein electrostriction: a possibility of elastic deformation of the α-hemolysin channel by the applied field. Eur Biophys J 34:997–1006CrossRefGoogle Scholar
  32. Krasnikov BF, Zorov DB, Antonenko YN, Zaspa AA, Kulikov IV, Kristal BS, Cooper AJL, Brown AM (2005) Comparative kinetic analysis reveals that inducer-specific ion release precedes the mitochondrial permeability transition. Biochim Biophys Acta 1708:375–392CrossRefGoogle Scholar
  33. La Rocca P, Shai Y, Sansom MS (1999) Peptide-bilayer interactions: simulations of dermaseptin B, an antimicrobial peptide. Biophys Chem 76:145–169CrossRefGoogle Scholar
  34. Lee M-T, Chen F-Y, Huang HW (2004) Energetics of pore formation induced by membrane active peptides. Biochemistry 43:3590–3599CrossRefGoogle Scholar
  35. Lequin O, Bruston F, Convert O, Chassaing G, Nicolas P (2003) Helical structure of dermaseptin B2 in a membrane-mimetic environment. Biochemistry 42:10311–10323CrossRefGoogle Scholar
  36. Lequin O, Ladram A, Chabbert L, Bruston F, Convert O, Vanhoye D, Chassaing G, Nicolas P, Amiche M (2006) Dermaseptin S9, an a-helical antimicrobial peptide with a hydrophobic core and cationic termini. Biochemistry 45:468–480CrossRefGoogle Scholar
  37. Lohner K, Blondelle SE (2005) Molecular mechanisms of membrane perturbation by antimicrobial peptides and the use of biophysical studies in the design of novel peptide antibiotics. Comb Chem High Throughput Screen 8:241–256CrossRefGoogle Scholar
  38. Magzoub M, Kilk K, Eriksson LEG, Langel U, Gräslund A (2001) Interaction and structure induction of cell-penetrating peptides in the presence of phospholipid vesicles. Biochim Biophys Acta 1512:77–89CrossRefGoogle Scholar
  39. Mai JC, Mi Z, Kim SH, Ng B, Robbins PD (2001) A proapoptotic peptide for the treatment of solid tumors. Cancer Res 61:7709–7712Google Scholar
  40. Mathew MK, Nagaraj R, Balaram P (1981) Alamethicin and synthetic peptide fragments as uncouplers of mitochondrial oxidative phosphorylation. Effect of chain length and charge. Biochem Biophys Res Commun 98:548–555CrossRefGoogle Scholar
  41. Montal M, Mueller P (1972) Formation of bimolecular membranes from monolayers and study of their electrical properties. Proc Natl Acad Sci USA 69:3561–3566CrossRefADSGoogle Scholar
  42. Mueller P, Rudin DO (1968) Action potentials induced in bimolecular lipid membranes. Nature 217:713–719CrossRefADSGoogle Scholar
  43. Oren Z, Shai Y (1998) Mode of action of linear amphipathic alpha-helical antimicrobial peptides. Biopolymers 47:451–463CrossRefGoogle Scholar
  44. Panchal RG, Smart ML, Bowser DN, Williams DA, Petrou S (2002) Pore-forming proteins and their application in biotechnology. Curr Pharm Biotechnol 3:99–115CrossRefGoogle Scholar
  45. Papo N, Shai Y (2003) Can we predict biological activity of antimicrobial peptides from their interactions with model phospholipid membranes. Peptides 24:1693–1703CrossRefGoogle Scholar
  46. Papo N, Shai Y (2005) Host defense peptides as new weapons in cancer treatment. Cell Mol Life Sci 62:784–790CrossRefGoogle Scholar
  47. Park CB, Yi KS, Matsuzaki K, Kim MS, Kim SC (2000) Structure–activity analysis of buforin II, a histone H2A-derived antimicrobial peptide: the proline hinge is responsible for the cell-penetrating ability of buforin II. Proc Natl Acad Sci USA 97:3245–8250Google Scholar
  48. Persson D, Thoren PE, Norden B (2001) Penetratin-induced aggregation and subsequent dissociation of negatively-charged phospholipid vesicles. FEBS Lett 505:307–312CrossRefGoogle Scholar
  49. Powers JP, Hancock REW (2003) The relationship between peptide structure and antibacterial activity. Peptides 24:1681–1691CrossRefGoogle Scholar
  50. Raimondo D, Andreotti G, Saint N, Amodeo P, Renzone G, Sanseverino M, Zocchi I, Molle G, Motta A, Scaloni A (2005) A folding-dependent mechanism of antimicrobial peptide resistance to degradation unveiled by solution structure of distinctin. Proc Natl Acad Sci USA 102:6309–6314CrossRefADSGoogle Scholar
  51. Rosenberger CM, Gallo RL, Finlay BB (2004) Interplay between antibacterial effectors: a macrophage antimicrobial peptide impairs Salmonella replication. Proc Natl Acad Sci USA 101:2422–2427 CrossRefADSGoogle Scholar
  52. Sahl HG, Pag U, Bouness S, Wagner S, Antcheva N, Toss A (2005) Mammalian defensins: structures and mechanism of action. J Leukoc Biol 77:466–475CrossRefGoogle Scholar
  53. Sheu MJT, Baldwin WW, Brunson KW (1985) Cytotoxicity of rabbit macrophage peptides MCP-1 and MCP-2 for mouse tumor cells. Antimicrob Agents Chemother 28:626–629Google Scholar
  54. Shaughnessy S, Nicholls P (1985) Control of respiration in sonicated cytochrome oxidase proteoliposomes by gated and ungated ionophores. Biochem Biophys Res Commun 128:1025–1030CrossRefGoogle Scholar
  55. Selsted ME, Ouelette AJ (2005) Mammalian defensins in the antimicrobial immune response. Nat Immunol 6:551–557CrossRefGoogle Scholar
  56. Seppet EK, Dhalla NS (1989) Characterization of Ca2+-stimulated ATPase in rat heart sarcolemma in the presence of dithiothreitol and alamethicin. Mol Cell Biochem 19:137–147CrossRefGoogle Scholar
  57. Wong JH, Ng TB (2005) Sesquin, a potent defensin-like antimicrobial peptide from ground beans with inhibitory activities toward tumor cells and HIV-1 transcriptase. Peptides 26:1120–1126CrossRefGoogle Scholar
  58. Wroblewski H, Burlot R, Johansson KE (1978) Solubilization of Spiroplasma citri cell membrane proteins with the anionic detergent sodium lauroyl-sarcosinate (Sarkosyl). Biochimie 60:389–398CrossRefGoogle Scholar
  59. Yang L, Harroun TA, Weiss TM, Ding L, Huang HW (2001) Barrel-stave or toroidal model? A case study on melittin pores. Biophys J 81:1475–1485Google Scholar
  60. Yonezawa A, Kuwahara J, Fujii N, Sugiura Y (1992) Binding of tachyplesin I to DNA revealed by footprinting analysis: significant contribution of secondary structure to DNA binding and implication for biological action. Biochemistry 31:2998–3004 CrossRefGoogle Scholar
  61. Yount NY, Yeaman MR (2004) Multidimensional signatures in antimicrobial peptides. Proc Natl Acad USA 101:7363–7368CrossRefADSGoogle Scholar
  62. Zemel A, Ben-Shaul A, May S (2005) Perturbation of a lipid membrane by amphipathic peptides and its role in pore formation. Eur Biophys J 34:230–242CrossRefGoogle Scholar
  63. Zhang L, Rozek A, Hancock RE (2001) Interaction of cationic antimicrobial peptides with model membranes. J Biol Chem 276:35714–35722CrossRefGoogle Scholar

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© EBSA 2006

Authors and Affiliations

  1. 1.Institut de Physiologie et de Biologie Cellulaires (Pôle Biologie Santé)UMR 6187 CNRS-Université de PoitiersPoitiersFrance

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