Abstract
Antimicrobial peptides are a large group of natural compounds which present promising properties for the pharmaceutical and food industries, such as broad-spectrum activity, potential for use as natural preservatives, and reduced propensity for development of bacterial resistance. Plantaricin 149 (Pln149), isolated from Lactobacillus plantarum NRIC 149, is an intrinsically disordered peptide with the ability to inhibit bacteria from the Listeria and Staphylococcus genera, and which is capable of promoting inhibition and disruption of yeast cells. In this study, the interactions of Pln149 with model membranes composed of zwitterionic and/or anionic phospholipids were investigated using a range of biophysical techniques, including isothermal titration calorimetry, surface tension measurements, synchrotron radiation circular dichroism spectroscopy, oriented circular dichroism spectroscopy, and optical microscopy, to elucidate these peptides’ mode of interactions and provide insight into their functional roles. In anionic model membranes, the binding of Pln149 to lipid bilayers is an endothermic process and induces a helical secondary structure in the peptide. The helices bind parallel to the surfaces of lipid bilayers and can promote vesicle disruption, depending on peptide concentration. Although Pln149 has relatively low affinity for zwitterionic liposomes, it is able to adsorb at their lipid interfaces, disturbing the lipid packing, assuming a similar parallel helix structure with a surface-bound orientation, and promoting an increase in the membrane surface area. Such findings can explain the intriguing inhibitory action of Pln149 in yeast cells whose cell membranes have a significant zwitterionic lipid composition.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abraham T, Lewis RN, Hodges RS, McElhaney RN (2005) Isothermal titration calorimetry studies of the binding of a rationally designed analogue of the antimicrobial peptide gramicidin S to phospholipid bilayer membranes. Biochemistry 44:2103–2112. https://doi.org/10.1021/bi048077d
Ageitos JM, Sánchez-Pérez A, Calo-Mata P, Villa TG (2017) Antimicrobial peptides (AMPs): ancient compounds that represent novel weapons in the fight against bacteria. Biochem Pharmacol 133:117–138. https://doi.org/10.1016/j.bcp.2016.09.018
Ambroggio EE, Separovic F, Bowie J, Fidelio GD (2004) Surface behaviour and peptide-lipid interactions of the antibiotic peptides, maculatin and citropin. Biochim Biophys Acta 1664:31–37. https://doi.org/10.1016/j.bbamem.2004.03.013
Angelova MI, Dimitrov DS (1986) Liposome electroformation. Faraday Discuss Chem Soc 81:303–311. https://doi.org/10.1039/DC9868100303
Bahar AA, Ren D (2013) Antimicrobial peptides. Pharmaceuticals (Basel) 6:1543–1575. https://doi.org/10.3390/ph6121543
Bechinger B, Gorr SU (2017) Antimicrobial peptides: mechanisms of action and resistance. J Dent Res 96:254–260. https://doi.org/10.1177/0022034516679973
Bozelli JC, Sasahara ET, Pinto MRS, Nakaie CR, Schreier S (2012) Effect of head group and curvature on binding of the antimicrobial peptide tritrpticin to lipid membranes. Chem Phys Lipids 165:365–373. https://doi.org/10.1016/j.chemphyslip.2011.12.005
Bürck J, Roth S, Wadhwani P, Afonin S, Kanithasen N, Strandberg E, Ulrich AS (2008) Conformation and membrane orientation of amphiphilic helical peptides by oriented circular dichroism. Biophys J 95:3872–3881. https://doi.org/10.1529/biophysj.108.136085
Bürck J, Roth S, Windisch D, Wadhwani P, Moss D, Ulrich AS (2015) UV-CD12: synchrotron radiation circular dichroism beamline at ANKA. J. Synchrotron Rad 22:844–852. https://doi.org/10.1107/S1600577515004476
Bürck J, Wadhwani P, Fanghänel S, Ulrich AS (2016) Oriented circular dichroism: a method to characterize membrane-active peptides in oriented lipid bilayers. Acc Chem Res 49:184–192. https://doi.org/10.1021/acs.accounts.5b00346
Cammers-Goodwin A, Allen TJ, Oslick SL, Mcclure KF, Lee JH, Kemp DS (1996) Mechanism of stabilization of helical conformations of polypeptides by water containing trifluoroethanol. J Am Chem Soc 118:3082–3090. https://doi.org/10.1021/ja952900z
Fernandez DI, Sani MA, Miles AJ, Wallace BA, Separovic F (2013) Membrane defects enhance the interaction of antimicrobial peptides, aurein 1.2 versus caerin 1.1. Biochim Biophys Acta 1828:1863–1872. https://doi.org/10.1016/j.bbamem.2013.03.010
Gordon YJ, Romanowski EG, McDermott AM (2005) A review of antimicrobial peptides and their therapeutic potential as anti-infective drugs. Curr Eye Res 30:505–515. https://doi.org/10.1080/02713680590968637
Hauge HH, Mantzilas D, Moll GN, Konings WN, Driessen AJ, Eijsink VG, Nissen-Meyer J (1998) Plantaricin A is an amphiphilic alpha-helical bacteriocin-like pheromone which exerts antimicrobial and pheromone activities through different mechanisms. Biochemistry 37(46):16026–16032. https://doi.org/10.1021/bi981532j
Henriksen JR, Andresen TL (2011) Thermodynamic profiling of peptide membrane interactions by isothermal titration calorimetry: a search for pores and micelles. Biophys J 101:100–109. https://doi.org/10.1016/j.bpj.2011.05.047
Iwamoto K, Hayakawa T, Murate M, Makino A, Ito K, Fujisawa T, Kobayashi T (2007) Curvature-dependent recognition of ethanolamine phospholipids by duramycin and cinnamycin. Biophys J 93:1608–1619. https://doi.org/10.1529/biophysj.106.101584
Kato T, Matsuda T, Ogawa E, Ogawa H, Kato H, Doi U, Nakamura R (1994) Plantaricin-149 a Bacteriocin Produced by Lactobacillus plantarum NRIC 149. J Ferment Bioeng 77:277–282. https://doi.org/10.1016/0922-338X(94)90234-8
Kenneth H, Rose AH (1972) Lipid Composition of Saccharomyces cereviseae as influenced by growth temperature. Biochim Biophys Acta Lipids Lipid Metab 260:639–653. https://doi.org/10.1016/0005-2760(72)90013-6
Koller D, Lohner K (2014) The role of spontaneous lipid curvature in the interaction of interfacially active peptides with membranes. Biochim Biophys Acta Biomembr 1838:2250–2259. https://doi.org/10.1016/j.bbamem.2014.05.013
Kristiansen PE, Fimland G, Mantzilas D, Nissen-Meyer J (2005) Structure and mode of action of the membrane-permeabilizing antimicrobial peptide pheromone Plantaricin A. J Biol Chem 280(24):22945–22950. https://doi.org/10.1074/jbc.M501620200
Kumagai PS, DeMarco R, Lopes JLS (2017) Advantages of synchrotron radiation circular dichroism spectroscopy to study intrinsically disordered proteins. Eur Biophys J 46:599–606. https://doi.org/10.1007/s00249-017-1202-1
Kumar P, Kizhakkedathu JN, Straus SK (2018) Antimicrobial peptides: diversity, mechanism of action and strategies to improve the activity and biocompatibility in vivo. Biomolecules 8:4. https://doi.org/10.3390/biom8010004
Lees JG, Smith BR, Wien F, Miles AJ, Wallace BA (2004) CDtool—an integrated software package for circular dichroism spectroscopic data processing, analysis, and archiving. Anal Biochem 332:285–289. https://doi.org/10.1016/j.ab.2004.06.002
Lindberg L, Santos AX, Riezman H, Olsson L (2013) Bettiga M (2013) Lipidomic profiling of Saccharomyces cerevisiae and Zygosaccharomyces bailii reveals critical changes in lipid composition in response to acetic acid stress. PLoS ONE 8(9):e73936. https://doi.org/10.1371/journal.pone.0073936 (eCollection)
Lopes JLS, Nobre TM, Siano A, Humpola V, Bossolan NRS, Zaniquelli MED, Tonarelli G, Beltramini LM (2009) Disruption of Saccharomyces cerevisiae by Plantaricin 149 and investigation of its mechanism of action with biomembrane model systems. Biochim Biophys Acta Biomembr 1788:2252–2258. https://doi.org/10.1016/j.bbamem.2009.06.026
Lopes JLS, Gómara MJ, Haro I, Tonarelli G, Beltramini LM (2013) Contribution of the Tyr-1 in Plantaricin 149a to disrupt phospholipid model membranes. Int J Mol Sci 14:12313–12328. https://doi.org/10.3390/ijms140612313
Lopes JLS, Miles AJ, Whitmore L, Wallace BA (2014) Distinct circular dichroism spectroscopic signatures of polyproline II and unordered secondary structures: applications in secondary structure analyses. Protein Sci 23:1765–1772. https://doi.org/10.1002/pro.2558
Marsh D (1996) Lateral pressure in membranes. Biochim Biophys Acta 1286:183–223. https://doi.org/10.1016/S0304-4157(96)00009-3
Marshall SH, Arenas G (2003) Antimicrobial peptides: a natural alternative to chemical antibiotics and a potential for applied biotechnology. Electron J Biotechnol 6:271–284. https://doi.org/10.2225/vol6-issue3-fulltext-1
Miles AJ, Wallace BA (2016) Circular dichroism spectroscopy of membrane proteins. Chem Soc Rev 45:4859–4872. https://doi.org/10.1039/C5CS00084J
Müller DM, Carrasco MS, Simonetta AC, Beltramini LM, Tonarelli GG (2007) A synthetic analog of Plantaricin 149 inhibiting food-borne pathogenic bacteria: evidence for α-helical conformation involved in bacteria-membrane interaction. J Peptide Sci 13:171–178. https://doi.org/10.1002/psc.828
Rai M, Pandit R, Gaikwad S, Kövics G (2016) Antimicrobial peptides as natural bio-preservative to enhance the shelf-life of food. J Food Sci Technol 53:3381–3394. https://doi.org/10.1007/s13197-016-2318-5
Sand SL, Haug TM, Nissen-Meyer J, Sand O (2007) The bacterial peptide pheromone Plantaricin A permeabilizes cancerous, but not normal, rat pituitary cells and differentiates between the outer and inner membrane leaflet. J. Membr Biol 216(2–3):61–71. https://doi.org/10.1007/s00232-007-9030-3
Sand SL, Oppegard C, Ohara S, Iijima T, Naderi S, Blomhoff HK, Nissen-Meyer J, Sand O (2010) Plantaricin A, a peptide pheromone produced by Lactobacillus plantarum, permeabilizes the cell membrane of both normal and cancerous lymphocytes and neuronal cells. Peptides 31(7):1237–1244. https://doi.org/10.1016/j.peptides.2010.04.010
Sand SL, Nissen-Meyer J, Sand O, Haug TM (2013) Plantaricin A, a cationic peptide produced by Lactobacillus plantarum, permeabilizes eukaryotic cell membranes by a mechanism dependent on negative surface charge linked to glycosylated membrane proteins. Biochim Biophys Acta Biomembr 1828(2), 249–259. https://doi.org/0.1016/j.bbamem.2012.11.001
Sani MA, Separovic F (2016) How membrane-active peptides get into lipid membranes. Acc Chem Res 49:1130–1138. https://doi.org/10.1021/acs.accounts.6b00074
Seelig J (1997) Titration calorimetry of lipid-peptide interactions. Biochim Biophys Acta 1331:103–116. https://doi.org/10.1016/S0304-4157(97)00002-6
Seelig J (2004) Thermodynamics of lipid-peptide interactions. Biochim Biophys Acta 1666:40–50. https://doi.org/10.1016/j.bbamem.2004.08.004
Shai Y (2002) Mode of action of membrane active antimicrobial peptides. Biopolymers 66(4):236–248. https://doi.org/10.1002/bip.10260
Singh VP (2018) Recent approaches in food bio-preservation—a review. Open Vet J 8:104–111. https://doi.org/10.4314/ovj.v8i1.16
Strandberg E, Tiltak D, Ehni S, Wadhwani P, Ulrich AS (2012) Lipid shape is a key factor for membrane interactions of amphipathic helical peptides. Biochim Biophys Acta Biomembr 1818:1764–1776. https://doi.org/10.1016/j.bbamem.2012.02.027
Tabaei SR, Rabe M, Zhdanov VP, Cho NJ, Höök F (2012) Single vesicle analysis reveals nanoscale membrane curvature selective pore formation in lipid membranes by an antiviral α-helical peptide. Nano Lett 12:5719–5725. https://doi.org/10.1021/nl3029637
Voievoda N, Schulthess T, Bechinger B, Seelig J (2015) Thermodynamics and biophysical analysis of the membrane-association of a histidine-rich peptide with efficient antimicrobial and transfection activities. J Phys Chem B 119:9678–9687. https://doi.org/10.1021/acs.jpcb.5b04543
Wallace BA (2009) Protein characterisation by synchrotron radiation circular dichroism spectroscopy. Q Rev Biophys 42:317–370. https://doi.org/10.1017/S003358351000003X
Wang S, Zeng X, Yang Q, Qiao S (2016) Antimicrobial peptides as potential alternatives to antibiotics in food animal industry. Int J Mol Sci 17:603–615. https://doi.org/10.3390/ijms17050603
Wieprecht T, Dathe M, Schumann M, Krause E, Beyermann M, Bienert M (1996) Conformational and functional study of magainin 2 in model membrane environments using the new approach of systematic double-d-amino acid replacement. Biochemistry 35:10844–10853. https://doi.org/10.1021/bi960362c
Wieprecht T, Apostolov O, Beyermann M, Seelig J (1999) Thermodynamics of the α-helix-coil transition of amphipathic peptides in a membrane environment: implications for the peptide- membrane binding equilibrium. J Mol Biol 294:785–794. https://doi.org/10.1006/jmbi.1999.3268
Wieprecht T, Apostolov O, Beyermann M, Seelig J (2000) Interaction of a mitochondrial presequence with lipid membranes: role of helix formation for membrane binding and perturbation. Biochemistry 39:15297–15305. https://doi.org/10.1021/bi001774v
Wimley WC, White SH (1996) Experimentally determined hydrophobicity scale for proteins at membrane interfaces. Nat Struct Biol 3:842–848. https://doi.org/10.1038/nsb1096-842
Zhao H, Sood R, Jutila A, Bose S, Fimland G, Nissen-Meyer J, Kinnunen PK (2006) Interaction of the antimicrobial peptide pheromone Plantaricin A with model membranes: implications for a novel mechanism of action. Biochim Biophys Acta 1758(9):1461–1474. https://doi.org/10.1016/j.bbamem.2006.03.037
Acknowledgements
We are grateful for the financial support of the following: a paired Biotechnology and Biological Sciences Research Council (BBSRC) Grant N012763/1/Sao Paulo Research Foundation (FAPESP) Grant 2015/50347-2 (to APUA and BAW), CNPq/PIBIC fellowship (to VKS), Grants 303513/2016-0 and 406429/2016-2 from CNPq-Brazil (to JLSL), FAPESP/CEPID Grant 2013/07600-3 (to LMB), FAPESP Grant 2018/19546-7 (to JLSL), Grant P02409 from the BBSRC (to BAW). APU, JLSL, LMB, and RI are recipients of research CNPq fellowships. Access to the AU-CD at ASTRID2 was supported by a beamtime grant (to PSK and JLSL). Access to beamline UV-CD12 of the Institute of Biological Interfaces (IBG2) and the Institute for Beam Physics and Technology (IBPT) storage ring, the Karlsruhe Research Accelerator (KARA) was enabled by grants from the Karlsruhe Institute of Technology.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Kumagai, P.S., Sousa, V.K., Donato, M. et al. Unveiling the binding and orientation of the antimicrobial peptide Plantaricin 149 in zwitterionic and negatively charged membranes. Eur Biophys J 48, 621–633 (2019). https://doi.org/10.1007/s00249-019-01387-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00249-019-01387-y