Abstract
Membrane-active peptides that demonstrate cell-penetrating, antimicrobial or cytotoxic functions are diverse in their amino acid sequences, but share common physicochemical features like short length, amphipathic conformation in membrane environment and high net charge. Nonspecific electrostatic interactions of basic peptide residues with anionic membrane lipids play a crucial role in the initial binding of such peptides to plasma membranes of bacterial and mammalian cells. At the same time, a number of membrane-active peptides functions when they are localized at high concentrations on the lipid membranes. Dissecting the role of electrostatics in this functional peptide conditions is important to understand why the majority of them bear high positive charge. We have studied interaction of EB1 cell-penetrating peptide (charge + 8) with model anionic membranes. The saturation of peptide binding to liposomes that comprises 5%, 10% and 25% of negatively charged lipids (POPC/POPG mixture) was observed. We have found that peptide recharges liposomes and its surface saturating concentration increases with the amount of anionic lipids in a membrane so as a surface charge (bound peptide plus anionic lipids). This observation may be explained with the Gouy–Chapman theory based model with addition of independent effective peptide charges for peptide–peptide and peptide–lipid interactions, as well as steric saturation term. Additionally, in certain conditions, membrane bound peptide leads to liposome aggregation. In some lipid-to-peptide ratio regions disaggregation follows that may indicate an additional slow equilibration process after fast initial peptide binding.
Similar content being viewed by others
References
Almeida PF (2014) Membrane-active peptides: binding, translocation, and flux in lipid vesicles. Biochim Biophys Acta (BBA) Biomembr 1838(9):2216–2227
Almeida PF, Ladokhin AS, White SH (2012) Hydrogen-bond energetics drive helix formation in membrane interfaces. Biochim Biophys Acta (BBA) Biomembr 1818(2):178–182
Arias M, Haney EF, Hilchie AL, Corcoran JA, Hyndman ME, Hancock RE, Vogel HJ (2020) Selective anticancer activity of synthetic peptides derived from the host defence peptide tritrpticin. Biochim Biophys Acta (BBA) Biomembr 1862(8):183228
Bechara C, Sagan S (2013) Cell-penetrating peptides: 20 years later, where do we stand? FEBS Lett 587(12):1693–1702
DeLano WL et al (2002) Pymol: an open-source molecular graphics tool. CCP4 Newslett Protein Crystallogr 40(1):82–92
Delaroche D, Aussedat B, Aubry S, Chassaing G, Burlina F, Clodic G, Bolbach G, Lavielle S, Sagan S (2007) Tracking a new cell-penetrating (w/r) nonapeptide, through an enzyme-stable mass spectrometry reporter tag. Anal Chem 79(5):1932–1938
Derossi D, Joliot AH, Chassaing G, Prochiantz A (1994) The third helix of the antennapedia homeodomain translocates through biological membranes. J Biol Chem 269(14):10444–10450
Durr UH, Gildenberg M, Ramamoorthy A (2012) The magic of bicelles lights up membrane protein structure. Chem Rev 112(11):6054–6074
Elmquist A, Lindgren M, Bartfai T, Langel Ü (2001) VE-cadherin-derived cell-penetrating peptide, pVEC, with carrier functions. Exp Cell Res 269(2):237–244
Epand RF, Maloy WL, Ramamoorthy A, Epand RM (2010) Probing the “charge cluster mechanism” in amphipathic helical cationic antimicrobial peptides. Biochemistry 49(19):4076–4084
Haney EF, Straus SK, Hancock RE (2019) Reassessing the host defense peptide landscape. Front Chem 7:43
Hellmann N, Schwarz G (1998) Peptide–liposome association. A critical examination with mastoparan-x. Biochim Biophys Acta (BBA) Biomembr 1369(2):267–277
Hennessey JP Jr, Johnson WC Jr (1982) Experimental errors and their effect on analyzing circular dichroism spectra of proteins. Anal Biochem 125(1):177–188
Islam MZ, Ariyama H, Alam JM, Yamazaki M (2014) Entry of cell-penetrating peptide transportan 10 into a single vesicle by translocating across lipid membrane and its induced pores. Biochemistry 53(2):386–396
Israelachvili JN (2011) Intermolecular and surface forces: revised third edition. Academic press, Cambridge
Jobin ML, Bonnafous P, Temsamani H, Dole F, Grélard A, Dufourc EJ, Alves ID (2013) The enhanced membrane interaction and perturbation of a cell penetrating peptide in the presence of anionic lipids: toward an understanding of its selectivity for cancer cells. Biochim Biophys Acta (BBA) Biomembr 1828(6):1457–1470
Klocek G, Schulthess T, Shai Y, Seelig J (2009) Thermodynamics of melittin binding to lipid bilayers. aggregation and pore formation. Biochemistry 48(12):2586–2596
Ladokhin AS, White SH (2001) Protein chemistry at membrane interfaces: non-additivity of electrostatic and hydrophobic interactions. J Mol Bio 309(3):543–552
Last NB, Miranker AD (2013) Common mechanism unites membrane poration by amyloid and antimicrobial peptides. Proc Natl Acad Sci 110(16):6382–6387
Lee DK, Brender JR, Sciacca MF, Krishnamoorthy J, Yu C, Ramamoorthy A (2013) Lipid composition-dependent membrane fragmentation and pore-forming mechanisms of membrane disruption by pexiganan (msi-78). Biochemistry 52(19):3254–3263
Lundberg P, El-Andaloussi S, Sütlü T, Johansson H, Langel Ü (2007) Delivery of short interfering RNA using endosomolytic cell-penetrating peptides. FASEB J 21(11):2664–2671
Murray D, Arbuzova A, Hangyás-Mihályné G, Gambhir A, Ben-Tal N, Honig B, McLaughlin S (1999) Electrostatic properties of membranes containing acidic lipids and adsorbed basic peptides: theory and experiment. Biophys J 77(6):3176–3188
Nguyen LT, Haney EF, Vogel HJ (2011) The expanding scope of antimicrobial peptide structures and their modes of action. Trends Biotechnol 29(9):464–472
Oehlke J, Scheller A, Wiesner B, Krause E, Beyermann M, Klauschenz E, Melzig M, Bienert M (1998) Cellular uptake of an \(\alpha \)-helical amphipathic model peptide with the potential to deliver polar compounds into the cell interior non-endocytically. Biochim Biophys Acta (BBA) Biomembr 1414(1–2):127–139
Peitzsch RM, Eisenberg M, Sharp KA, McLaughlin S (1995) Calculations of the electrostatic potential adjacent to model phospholipid bilayers. Biophys J 68(3):729–738
Persson D, Thorén PE, Nordén B (2001) Penetratin-induced aggregation and subsequent dissociation of negatively charged phospholipid vesicles. FEBS Lett 505(2):307–312
Sauder R, Seelig J, Ziegler A (2011) Thermodynamics of Lipid Interactions with Cell-Penetrating Peptides. In: Langel Ü (ed) Cell-Penetrating Peptides. Methods in Molecular Biology (Methods and Protocols), vol 683. (Humana Press). https://doi.org/10.1007/978-1-60761-919-2_10
Seelig J (2004) Thermodynamics of lipid–peptide interactions. Biochim Biophys Acta (BBA) Biomembr 1666(1):40–50
Splith K, Neundorf I (2011) Antimicrobial peptides with cell-penetrating peptide properties and vice versa. Eur Biophys J 40(4):387–397
Stankowski S (1991) Surface charging by large multivalent molecules. Extending the standard Gouy–Chapman treatment. Biophys J 60(2):341–351
Tamba Y, Yamazaki M (2009) Magainin 2-induced pore formation in the lipid membranes depends on its concentration in the membrane interface. J Phys Chem B 113(14):4846–4852
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(37):12018–12026
Thorén PE, Persson D, Lincoln P, Nordén B (2005) Membrane destabilizing properties of cell-penetrating peptides. Biophys Chem 114(2–3):169–179
Vives E, Brodin P, Lebleu B (1997) A truncated HIV-1 tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. J Biol Chem 272(25):16010–16017
Wadhwani P, Epand R, Heidenreich N, Bürck J, Ulrich A, Epand R (2012) Membrane-active peptides and the clustering of anionic lipids. Biophys J 103(2):265–274
White SH, Wimley WC (1999) Membrane protein folding and stability: physical principles. Annu Rev Biophys Biomol Struct 28(1):319–365
Wieprecht T, Beyermann M, Seelig J (1999) Binding of antibacterial magainin peptides to electrically neutral membranes: thermodynamics and structure. Biochemistry 38(32):10377–10387
Wimley WC, Hristova K (2011) Antimicrobial peptides: successes, challenges and unanswered questions. J Membr Biol 239(1–2):27–34
Acknowledgements
We are grateful to academician Michael Dubina for organizational and inspirational support of this study. The work was supported by RFBR grant no. 18-34-00992
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Svirina, A., Terterov, I. Electrostatic effects in saturation of membrane binding of cationic cell-penetrating peptide. Eur Biophys J 50, 15–23 (2021). https://doi.org/10.1007/s00249-020-01476-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00249-020-01476-3