Skip to main content

Action of antimicrobial peptides and cell-penetrating peptides on membrane potential revealed by the single GUV method

Abstract

Membrane potential plays various key roles in live bacterial and eukaryotic cells. So far, the effects of membrane potential on action of antimicrobial peptides (AMPs) and cell-penetrating peptides (CPPs) have been examined using cells and small lipid vesicles. However, due to the technical drawbacks of these experiments, the effect of membrane potential on the actions of AMPs and CPPs and the elementary processes of interactions of these peptides with cell membranes and vesicle membranes are not well understood. In this short review, we summarize the results of the effect of membrane potential on the action of an AMP, lactoferricin B (LfcinB), and a CPP, transportan 10 (TP10), in vesicle membranes revealed by the single giant unilamellar vesicle (GUV) method. Parts of the actions and their elementary steps of AMPs and CPPs interacting vesicle membranes under membrane potential are clearly revealed using the single GUV method. The experimental methods and their analysis described here can be used to elucidate the effects of membrane potential on various activities of peptides such as AMPs, CPPs, and proteins. Moreover, GUVs with membrane potential are more suitable as a model of cells or artificial cells, as well as GUVs containing small vesicles.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3

References

  1. Alberts B, Johnson A, Lewis J, Morgan D, Raff M, Roberts K, Walter P (2015) Molecular biology of the cell, 6th edn. Garland Science, New York

    Google Scholar 

  2. Andersen OS (1983) Ion movement through gramicidin a channels: interfacial polarization effects on single-channel current measurements. Biophys J 41:135–146

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Bechara C, Sagan S (2013) Cell-penetrating peptides: 20 years later, where do we stand? FEBS Lett 587:1693–1702

    CAS  PubMed  Google Scholar 

  4. Bechniger B, Zasloff M, Opella SJ (1993) Structure and orientation of the antibiotic peptide magainin in membranes by solid-state nuclear magnetic resonance spectroscopy. Protein Sci 2:2077–2084

    Google Scholar 

  5. Bellamy W, Takase M, Yamauchi K, Wakabayashi H, Kawase K, Tomita M (1992) Identification of the bacterial domain of lactoferrin. Biochim Biophys Acta 1121:130–136

    CAS  PubMed  Google Scholar 

  6. Blackiston DJ, McLaughlin KA, Levin M (2009) Bioelectric controls of cell proliferation. Cell Cycle 8:3527–3536

    CAS  PubMed  Google Scholar 

  7. Bobone S, Stella L (2019) Selectivity of antimicrobial peptides; a complex interplay of multiple equilibria. In: Matsuzaki K (ed) Antimicrobial peptides: basic for clinical application. Springer, New York, pp 17–32

    Google Scholar 

  8. Christensen B, Fink J, Merrifield RB, Mauzerall D (1988) Channel-forming properties of cecropins and related model compounds incorporated into planar lipid membranes. Proc Natl Acad Sci U S A 85:5072–5076

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Daniels CJ, Bole DG, Quay SC, Oxender DL (1981) Role for membrane potential in the secretion of protein into the periplasm of Escherichia coli. Proc Natl Acad Sci U S A 78:5396–5400

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Dubey GP, Ben-Yehuda S (2011) Intercellular nanotubes mediate bacterial communication. Cell 144:590–600

    CAS  PubMed  Google Scholar 

  11. EL-Andaloussi S, Johansson H, Magnusdottir A, Järver P, Lundberg P, Langel Ű (2005) TP10, a delivery vector for decoy oligonucleotides targeting the Myc protein. J Control Release 110:189–201

    CAS  PubMed  Google Scholar 

  12. Fanghänel S, Wadhwani P, Strandberg E, Verdurmen WPR, Bűeck J, Ehni S, Mykhailiuk PK, Afonin S, Gerthsen D, Komarov IV, Brock R, Ulrich AS (2014) Structure analysis and conformational transitions of the cell penetrating peptide transportan 10 in the membrane-bound state. PLoS One 9:e99653

    PubMed  PubMed Central  Google Scholar 

  13. Felle H, Porter JS, Slayman CL, Kaback HR (1980) Quantitative measurements of membrane potential in Escherichia coli. Biochemistry 19:3585–3590

    CAS  PubMed  Google Scholar 

  14. Finkelstein A, Andersen OS (1981) The gramicidin a channel: a review of its permeability characteristics with special reference to the single-file aspect of transport. J Membr Biol 59:155–171

    CAS  PubMed  Google Scholar 

  15. Futaki S, Suzuki T, Ohashi W, Yagami T, Tanaka S, Ueda K, Sugiura Y (2001) Arginine-rich peptides. An abundant source of membrane-permeable peptides having potential as carriers for intracellular protein delivery. J Biol Chem 276:5836–5840

    CAS  PubMed  Google Scholar 

  16. Gesell J, Zasloff M, Opella SJ (1997) Two-dimensional 1H NMR experiments show that the 23-residue magainin antibiotic peptide is an α-helix in dodecylphosphocholine micelles, sodium dodecylsulfate micelles, and trifluoroethanol/water solution. J Biomol NMR 9:127–135

    CAS  PubMed  Google Scholar 

  17. Hancock REW, Hans-Georg S (2006) Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nat Biotechnol 24:1551–1557

    CAS  PubMed  Google Scholar 

  18. Hancock REW, Rozek A (2002) Role of membranes in the activities of antimicrobial cationic peptides. FEMS Microbiol Lett 206:143–149

    CAS  PubMed  Google Scholar 

  19. Hasan M, Yamazaki M (2019) Elementary processes and mechanisms of interactions of antimicrobial peptides with membranes - single GUV studies. In: Matsuzaki K (ed) Antimicrobial peptides: basic for clinical application. Springer, New York, pp 17–32

    Google Scholar 

  20. Hasan M, Karal MAS, Levadnyy V, Yamazaki M (2018) Mechanism of initial stage of pore formation induced by antimicrobial peptide magainin 2. Langmuir 34:3349–3362

    CAS  PubMed  Google Scholar 

  21. Hasan M, Moghal MMR, Saha SK, Yamazaki M (2019) The role of membrane tension in the action of antimicrobial peptides and cell-penetrating peptides in biomembranes. Biophys Rev 11:431–448

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Henriques ST, Costa J, Castanho MARB (2005) Translocation of β-galactosidase mediated by the cell-penetrating peptide Pep-1 into lipid vesicles and human HeLa cells is driven by membrane electrostatic potential. Biochemistry 44:10189–10198

    PubMed  Google Scholar 

  23. Hille B (1992) Ionic channels of excitable membranes, 2nd edn. Sinauer Association Inc, Massachusetts

    Google Scholar 

  24. Hossain F, Moghal MMR, Islam MZ, Moniruzzaman M, Yamazaki M (2019) Membrane potential is vital for rapid permeabilization of plasma membranes and lipid bilayers by the antimicrobial peptide lactoferricin B. J Biol Chem 294:10449–10462

    CAS  PubMed  Google Scholar 

  25. Hwang PM, Zhou N, Shan X, Arrosmith CH, Vogel HJ (1998) Three-dimensional solution structure of lactoferricin B, an antimicrobial peptide derived from bovine lactoferrin. Biochemistry 37:4288–4298

    CAS  PubMed  Google Scholar 

  26. Ishihara Y, Shimamoto N (2006) Involvement of endonuclease G in nucleosomal DNA fragmentation under sustained endogenous oxidative stress. J Biol Chem 281:6726–6733

    CAS  PubMed  Google Scholar 

  27. Islam MZ, Ariyama H, Alam JM, Yamazaki M (2014a) Entry of cell-penetrating peptide transportan 10 into a single vesicle by translocating across lipid membrane and its induced pores. Biochemistry 53:386–396

    CAS  PubMed  Google Scholar 

  28. Islam MZ, Alam JM, Tamba Y, Karal MAS, Yamazaki M (2014b) The single GUV method for revealing the functions of antimicrobial, pore-forming toxin, and cell-penetrating peptides or proteins. Phys Chem Chem Phys 16:15752–15767

    CAS  PubMed  Google Scholar 

  29. Islam MZ, Sharmin S, Levadnyy V, Shibly SUA, Yamazaki M (2017) Effects of mechanical properties of lipid bilayers on entry of cell-penetrating peptides into single vesicles. Langmuir 33:2433–2443

    CAS  PubMed  Google Scholar 

  30. Islam MZ, Sharmin S, Moniruzzaman M, Yamazaki M (2018) Elementary processes for the entry of cell-penetrating peptides into lipid bilayer vesicles and bacterial cells. Appl Microbiol Biotechnol 102:3879–3892

    CAS  PubMed  Google Scholar 

  31. Kagan BL, Selsted ME, Ganz T, Lehrer R (1990) Antimicrobial defensin peptides from voltage-dependent ion-permeable channels in planar lipid bilayer membranes. Proc Natl Acad Sci U S A 87:210–214

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Karal MAS, Alam JM, Takahashi T, Levadny V, Yamazaki M (2015) Stretch-activated pore of antimicrobial peptide Magainin 2. Langmuir 31:3391–3401

    CAS  PubMed  Google Scholar 

  33. Kralj JM, Hochbaum DR, Douglass AD, Cohen AE (2011) Electrical spiking in Escherichia coli probed with a fluorescent voltage-indicating protein. Science 333:345–348

    CAS  PubMed  Google Scholar 

  34. Kuwata H, Yip TT, Tomita M, Hutchens TW (1998) Direct evidence of the generation in human stomach of an antimicrobial peptide domain (lactoferricin) from ingested lactoferrin. Biochim Biophys Acta 1429:129–141

    CAS  PubMed  Google Scholar 

  35. Lin C-C, Bachmann M, Bachler S, Venkatesan K, Dittrich PS (2018) Tunable membrane potential reconstituted in giant vesicles promotes permeation of cationic peptides at nanomolar concentrations. ACS Appl Mater Interfaces 10:41909–41916

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Madani F, Lindberg S, Langel Ű, Futaki S, Gräslund A (2011) Mechanisms of cellular uptake of cell-penetrating peptides. J Biophysics 414729

  37. Martinac B, Buechner M, Delcour AH, Adler J, Kung C (1987) Pressure-sensitive ion channel in Escherichia coli. Proc Natl Acad Sci U S A 84:2297–2301

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Melo MN, Ferre R, Castanho ARB (2009) Antimicrobial peptides: linking partition, activity and high membrane-bound concentrations. Nat Rev Microbiol 8:1–5

    Google Scholar 

  39. Moghal MMR, Islam MZ, Sharmin S, Levadnyy V, Moniruzzaman M, Yamazaki M (2018) Continuous detection of entry of cell-penetrating peptide Transportan 10 into single vesicles. Chem Phys Lipids 212:120–129

    CAS  PubMed  Google Scholar 

  40. Moghal MMR, Islam MZ, Hossain F, Saha SK, Yamazaki M (2020) Role of membrane potential on entry of cell-penetrating peptide transportan 10 into single vesicles. Biophys J 118:57–60

    CAS  PubMed  Google Scholar 

  41. Moniruzzaman M, Alam JM, Dohra H, Yamazaki M (2015) Antimicrobial peptide lactoferricin B-induced rapid leakage of internal contents from single giant unilamellar vesicles. Biochemistry 54:5802–5814

    CAS  PubMed  Google Scholar 

  42. Moniruzzaman M, Islam MZ, Sharmin S, Dohra H, Yamazaki M (2017) Entry of a six-residue antimicrobial peptide derived from Lactoferricin B into single vesicles and Escherichia coli cells without damaging their membranes. Biochemistry 56:4419–4431

    CAS  PubMed  Google Scholar 

  43. Pasupleti M, Schmidtchen A, Malmsten M (2012) Antimicrobial peptides: key components of the innate immune system. Crit Rev Biotechnol 32:143–171

    Google Scholar 

  44. Pautot S, Frisken BJ, Weitz DA (2003) Production of unilamellar vesicles using an inverted emulsion. Langmuir 19:2870–2879

    CAS  Google Scholar 

  45. Pelkmans L, Helenius A (2003) Insider information: what viruses tell us about endocytosis. Curr Opin Cell Biol 15:414–422

    CAS  PubMed  Google Scholar 

  46. Propheter DC, Chara AL, Harris TA, Ruhn KA, Hooper LV (2017) Resistin-like molecule β is a bactericidal protein that promotes spatial segregation of the microbiota and the colonic epithelium. Proc Natl Acad Sci U S A 114:11027–11033

    CAS  PubMed  PubMed Central  Google Scholar 

  47. Richardson BD, Saha K, Krout D, Cabrera E, Felts B, Henry LK, Swant J, Zou M-F, Newman AH, Khoshbouei H (2015) Membrane potential shapes regulation of dopamine transporter trafficking at the plasma membrane. Nat Commun 7:10423

    Google Scholar 

  48. Rothbard JB, Jessop TC, Lewis RS, Murray BA, Wender PA (2004) Role of membrane potential and hydrogen bonding in the mechanism of translocation of guanidinium-rich peptides into cells. J Am Chem Soc 126:9506–9507

    CAS  PubMed  Google Scholar 

  49. Rothbard JB, Jessop TC, Wender PA (2005) Adaptive translocation: the role of hydrogen bonding and membrane potential in the uptake of guanidinium-rich transporters into cells. Adv Drug Deliv Rev 57:495–504

    CAS  PubMed  Google Scholar 

  50. Ruthe H-J, Adler J (1985) Fusion of bacterial spheroplasts by electric fields. BBA-Biomembranes 819:105–113

    CAS  PubMed  Google Scholar 

  51. Shapiro HM (1994) Cell membrane potential analysis. Methods Cell Biol 41:121–133

    CAS  PubMed  Google Scholar 

  52. Sharmin S, Islam MZ, Karal MAS, Shibly SUA, Dohra H, Yamazaki M (2016) Effects of lipid composition on the entry of cell-penetrating peptide oligoarginine into single vesicles. Biochemistry 55:4154–4165

    CAS  PubMed  Google Scholar 

  53. Shibly SUA, Ghatak C, Karal MAS, Moniruzzaman MM. Yamazaki M (2016) Experimental estimation of membrane tension induced by osmotic pressure. Biophys J 111:2190–2201

  54. Sochacki KA, Barns KJ, Bucki R, Weisshaar JC (2011) Real-time attack on single Escherichia coli cells by the human antimicrobial peptide LL-37. Proc Natl Acad Sci U S A 108:E77–E81

    PubMed  PubMed Central  Google Scholar 

  55. Soomets U, Lindgren M, Gallet X, Pooga M, Hällbrink M, Elmquist A, Balaspiri L, Zorko M, Pooga M, Brasseur R, Langel Ű (2000) Deletion analogues of transportan. Biochim Biophys Acta 1467:165–176

    CAS  PubMed  Google Scholar 

  56. Sperelakis N (2012) Cell physiology source book, Essentials of membrane biophysics, 4th edn. Academic press, London

    Google Scholar 

  57. Stanzl EG, Trantow BM, Vargas JR, Wender PA (2013) Fifteen years of cell-penetrating, guanidinium-rich molecular transporters: basic science, research tools, and clinical applications. Acc Chem Res 46:2944–2954

    CAS  PubMed  Google Scholar 

  58. Strahl H, Hamoen LW (2010) Membrane potential is important for bacterial cell division. Proc Natl Acad Sci U S A 107:12281–12286

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Stratford JP, Edwards CLA, Ghanshyam MJ, Malyshev D, Delise MA, Hayashi Y, Asally M (2019) Electrically induced bacterial membrane-potential dynamics correspond to cellular proliferation capacity. Proc Natl Acad Sci U S A 116:9552–9557

    CAS  PubMed  PubMed Central  Google Scholar 

  60. Sun Y, Sun T-L, Huang HW (2016) Mode of action of antimicrobial peptides on E. coli spheroplasts. Biophys J 111:132–139

    CAS  PubMed  PubMed Central  Google Scholar 

  61. Sunderacruz S, Levin M, Kaplan DL (2009) Role of membrane potential in the regulation of cell proliferation and differentiation. Stem Cell Rev Rep 5:231–246

    Google Scholar 

  62. Tamba Y, Yamazaki M (2005) Single giant unilamellar vesicle method reveals effect of antimicrobial peptide, magainin 2, on membrane permeability. Biochemistry 44:15823–15833

    CAS  PubMed  Google Scholar 

  63. Tamba Y, Yamazaki M (2009) Magainin 2-induced pore formation in membrane depends on its concentration in membrane interface. J Phys Chem B 113:4846–4852

    CAS  PubMed  Google Scholar 

  64. Tamba Y, Ohba S, Kubota M, Yoshioka H, Yoshioka H, Yamazaki M (2007) Single GUV method reveals interaction of tea catechin (−)-epigallocatechin gallate with lipid membranes. Biophys J 92:3178–3194

    CAS  PubMed  PubMed Central  Google Scholar 

  65. Tamba Y, Ariyama H, Levadny V, Yamazaki M (2010) Kinetic pathway of antimicrobial peptide magainin 2-induced pore formation in lipid membranes. J Phys Chem B 114:12018–12026

    CAS  PubMed  Google Scholar 

  66. Tamba Y, Terashima H, Yamazaki M (2011) A membrane filtering method for the purification of giant unilamellar vesicles. Chem Phys Lipids 164:351–358

    CAS  PubMed  Google Scholar 

  67. Terrone D, Sang SLW, Roudaia L, Silvius JR (2003) Penetratin and related cell-penetrating cationic peptides can translocate across lipid bilayers in the presence of a transbilayer potential. Biochemistry 42:13787–13799

    CAS  PubMed  Google Scholar 

  68. Walde P, Cosentino K, Engel H, Stano P (2010) Giant vesicles: preparations and applications. ChemBioChem 11:848–865

    CAS  PubMed  Google Scholar 

  69. Wallbrecher R, Ackels T, Olea RA, Klein MJ, Caillon L, Schiller J, Bovée-Geurts PH, van Kuppevelt TH, Ulrich AS, Spehr M, Adjobo-Hermans MJW, Brock R (2017) Membrane permeation of arginine-rich cell-penetrating peptides independent of transmembrane potential as a function of lipid composition and membrane fluidity. J Control Release 256:68–78

    CAS  PubMed  Google Scholar 

  70. Wei L, LaBouyer MA, Darling LEO, Elmore DE (2016) Bacterial spheroplasts as a model for visualizing membrane translocation of antimicrobial peptides. Antimicrob Agents Chemother 60:6350–6352

    CAS  PubMed  PubMed Central  Google Scholar 

  71. Wu M, Maier E, Benz R, Hancock REW (1999) Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochemistry 38:7235–7242

    CAS  PubMed  Google Scholar 

  72. Yamazaki M (2008) The single GUV method to reveal elementary processes of leakage of internal contents from liposomes induced by antimicrobial substances. Adv Planar Lipid Bilayers Liposomes 7:121–142

    CAS  Google Scholar 

  73. Yeaman MR, Yount NY (2003) Mechanisms of antimicrobial peptide action and resistance. Pharmacol Rev 55:27–55

    CAS  PubMed  Google Scholar 

  74. Yeaman MR, Bayer AS, Koo S-P, Foss W, Sullam PM (1998) Platelet microbicidal proteins and neutrophil defensing disrupt the Staphylococcus aureus cytoplasmic membrane by distinct mechanism of action. J Clin Invest 101:178–187

    CAS  PubMed  PubMed Central  Google Scholar 

  75. Zasloff M (1987) Magainins, a class of antimicrobial peptides from Xenopus skin: isolation, characterization of two active forms, and partial cDNA sequence of a precursor. Proc Natl Acad Sci U S A 84:5449–5453

    CAS  PubMed  PubMed Central  Google Scholar 

  76. Zasloff M (2002) Antimicrobial peptides of multicellular organisms. Nature 415:389–395

    CAS  PubMed  Google Scholar 

  77. Zhang X, Jin Y, Plummer MR, Pooyan S, Gunaseelan S, Sinko PJ (2009) Endocytosis and membrane potential are required for Hela cell uptake of R.I.-CKTat9, a retro-inverso tat cell penetrating peptide. Mol Pham 6:836–848

    CAS  Google Scholar 

  78. Zhou Y, Wong C-O, Cho K-J, van der Hoeven D, Liang H, Thakur DP, Luo J, Babic M, Zinsmaier KE, Zhu MX, Hu H, Venkatachalam K, Hancock JF (2015) Membrane potential modulates plasma membrane phospholipid dynamics and K-Ras signaling. Science 349:873–876

    CAS  PubMed  PubMed Central  Google Scholar 

  79. Zorko M, Langel Ű (2005) Cell-penetrating peptides: mechanism and kinetics of cargo delivery. Adv Drug Deliv Rev 57:529–545

    CAS  PubMed  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Masahito Yamazaki.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Moghal, M.M.R., Hossain, F. & Yamazaki, M. Action of antimicrobial peptides and cell-penetrating peptides on membrane potential revealed by the single GUV method. Biophys Rev 12, 339–348 (2020). https://doi.org/10.1007/s12551-020-00662-z

Download citation

Keywords

  • Single giant unilamellar vesicles
  • Antimicrobial peptides
  • Cell-penetrating peptides
  • Membrane potential
  • Artificial cells
  • Bio-imaging