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Location of Peptide-Induced Submicron Discontinuities in the Membranes of Vesicles Using ImageJ

Abstract

Cell penetrating peptide transportan 10 and antimicrobial peptide melittin formed submicron pores in the lipid membranes of vesicles which are explained by the leakage of water-soluble fluorescent probes from the inside of vesicles to the outside. It is hypothesized that these submicron pores induce submicron discontinuities in the membranes. Considering this hypothesis, a technique has developed to locate the submicron discontinuities in the membranes of giant unilamellar vesicles (GUVs) using ImageJ. In this technique, at first the edges of membrane of a ‘single GUV’ are detected and then these edges are used to locate the submicron discontinuities. Two continuous rings are observed after applying the ImageJ in GUVs which indicated the edges of membrane. In contrast, the submicron discontinuations are detected at the edges of transportan 10 and melittin induced pore formed membranes. This investigation might be helpful for the elucidation of mechanism of the peptide-induced pore formation in the membranes of vesicles.

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References

  1. 1.

    Rawicz W, Olbrich KC, McIntosh T, Needham D, Evans E (2000) Effect of chain length and unsaturation on elasticity of lipid bilayers. Biophys J 79:328–339. https://doi.org/10.1016/S0006-3495(00)76295-3

  2. 2.

    Lee M-T, Sun T-L, Hung W-C, Huang HW (2013) Process of inducing pores in membranes by melittin. Proc Natl Acad Sci 110:14243–14248. https://doi.org/10.1073/pnas.1307010110

    Article  PubMed  Google Scholar 

  3. 3.

    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 53:386–396. https://doi.org/10.1021/bi401406p

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Islam MZ, Alam JM, Tamba Y, Karal MAS, Yamazaki M (2014) 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. https://doi.org/10.1039/c4cp00717d

  5. 5.

    Karal MAS, Alam JM, Takahashi T, Levadny V, Yamazaki M (2015) Stretch-activated pore of the antimicrobial peptide, magainin 2. Langmuir 31:3391–3401. https://doi.org/10.1021/la503318z

  6. 6.

    Karal MAS, Levadnyy V, Tsuboi T-A,  Belaya M, Yamazaki M (2015) Electrostatic interaction effects on tension-induced pore formation in lipid membranes. Phys Rev E 92:012708. https://doi.org/10.1103/PhysRevE.92.012708

  7. 7.

    Karal MAS, Ahmed M, Levadny V, Belaya M, Ahamed MK, Rahman M, Shakil MM (2020) Electrostatic interaction effects on the size distribution of self-assembled giant unilamellar vesicles. Phys Rev E 101:012404. https://doi.org/10.1103/PhysRevE.101.012404

  8. 8.

    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. https://doi.org/10.1021/acs.biochem.6b00189

  9. 9.

    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. https://doi.org/10.1021/acs.langmuir.7b04219

  10. 10.

    Karal MAS, Islam MK, Mahbub ZB (2020) Study of molecular transport through a single nanopore in the membrane of a giant unilamellar vesicle using COMSOL simulation. Eur Biophys J 49:59–69. https://doi.org/10.1007/s00249-019-01412-0

  11. 11.

    Sandre O, Moreaux L, Brochard-Wyart F (1999) Dynamics of transient pores in stretched vesicles. Proc Natl Acad Sci 96:10591–10596. https://doi.org/10.1073/pnas.96.19.10591

  12. 12.

    Riske KA, Dimova R (2005) Electro-deformation and poration of giant vesicles viewed with high temporal resolution. Biophys J 88:1143–1155. https://doi.org/10.1529/biophysj.104.050310

  13. 13.

    Zhang D, Karki AB, Rutman D, Young DP, Wang A, Cocke D, Ho TH, Guo Z (2009) Electrospun polyacrylonitrile nanocomposite fibers reinforced with Fe3O4 nanoparticles: fabrication and property analysis. Polymer 50:4189–4198. https://doi.org/10.1016/j.polymer.2009.06.062

  14. 14.

    Shibly SUA, Ghatak C, Karal MAS, Moniruzzaman M, Yamazaki M (2016) Experimental estimation of membrane tension induced by osmotic pressure. Biophys J 111:2190–2201. https://doi.org/10.1016/j.bpj.2016.09.043

  15. 15.

    Karal MAS, Ahamed MK, Rahman M, Ahmed M,  Shakil MM, Rabbani KS (2019) Effects of electrically-induced constant tension on giant unilamellar vesicles using irreversible electroporation. Eur Biophys J 48:731–741. https://doi.org/10.1007/s00249-019-01398-9

  16. 16.

    Sezgin E (2017) Super-resolution optical microscopy for studying membrane structure and dynamics. J Phys Condens Matter 29:273001. https://doi.org/10.1088/1361-648X/aa7185

    Article  PubMed  PubMed Central  Google Scholar 

  17. 17.

    Portet T, Dimova R (2010) A new method for measuring edge tensions and stability of lipid bilayers: effect of membrane composition. Biophys J 99:3264–3273. https://doi.org/10.1016/j.bpj.2010.09.032

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Fuertes G, Giménez D, Esteban-Martín S et al (2011) A lipocentric view of peptide-induced pores. Eur Biophys J 40:399–415. https://doi.org/10.1007/s00249-011-0693-4

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Ferreira T, Rasband W (2011) The imageJ user guide (ImageJ 1.44). online at http://imagej.nih.gov/ij/docs/guide/

  20. 20.

    Eliceiri KW, Rueden C (2005) Tools for visualizing multidimensional images from living specimens. Photochem Photobiol 81:1116–1122. https://doi.org/10.1562/2004-11-22-IR-377

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to imageJ: 25 years of image analysis. Nat Methods 9:671–675. https://doi.org/10.1038/nmeth.2089

  22. 22.

    Karal MAS, Levadnyy V, Yamazaki M (2016) Analysis of constant tension-induced rupture of lipid membranes using activation energy. Phys Chem Chem Phys 18:13487–13495. https://doi.org/10.1039/C6CP01184E

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Karal MAS, Rahman M, Ahamed MK, Shibly SUA, Ahmed M, Shakil MM (2019) Low cost non-electromechanical technique for the purification of giant unilamellar vesicles. Eur Biophys J 48:349–359. https://doi.org/10.1007/s00249-019-01363-6

  24. 24.

    Karal MAS, Yamazaki M (2015) Communication: activation energy of tension-induced pore formation in lipid membranes. J Chem Phys 143:081103. https://doi.org/10.1063/1.4930108

  25. 25.

    Tamba Y, Terashima H, Yamazaki M (2011) A membrane filtering method for the purification of giant unilamellar vesicles. Chem Phys Lipids 164:351–358. https://doi.org/10.1016/j.chemphyslip.2011.04.003

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Klette R (2014) Concise computer vision (An introduction into theory and algorithms). Springer London. https://doi.org/10.1007/978-1-4471-6320-6

  27. 27.

    Bihler M (2014) Overview of edge and line detection on FPGAs and implementation of an FPGA-based edge detector (Technical Report). https://doi.org/10.13140/RG.2.1.2305.7442

  28. 28.

    Bleicken S, Wagner C, García-Sáez AJ (2013) Mechanistic differences in the membrane activity of bax and Bcl-xL correlate with their opposing roles in apoptosis. Biophys J 104:421–431. https://doi.org/10.1016/j.bpj.2012.12.010

  29. 29.

    Bleicken S, Landeta O, Landajuela A,  Basañez G, García-Sáez AJ (2013) Proapoptotic bax and bak proteins form stable protein-permeable pores of tunable size. J Biol Chem 288:33241–33252. https://doi.org/10.1074/jbc.M113.512087

  30. 30.

    Jacobson K, Ishihara A, Inman R (1987) Lateral diffusion of proteins in membranes. Annu Rev Physiol 49:163–175. https://doi.org/10.1146/annurev.ph.49.030187.001115

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Lipowsky R, Sackmann E (1995) Structure and dynamics of membranes: I. From cells to vesicles / II. Generic and specific interactions. Elsevier, North Holland

  32. 32.

    Karal MAS, Ahammed S, Levadny V, Belaya M, Ahamed MK, Ahmed M, Mahbub ZB, Ullah AKMA (2020) Deformation and poration of giant unilamellar vesicles induced by anionic nanoparticles. Chem Phys Lipids 230:104916. https://doi.org/10.1016/j.chemphyslip.2020.104916

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Acknowledgements

This work was supported partly by the Grants from Ministry of Science and Technology, Ministry of Education, ICT Division (Ministry of Posts, Telecommunications and Information Technology) and CASR-BUET, Bangladesh to Mohammad Abu Sayem Karal.

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Correspondence to Mohammad Abu Sayem Karal.

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Karal, M.A.S., Ahamed, M.K., Ahmed, M. et al. Location of Peptide-Induced Submicron Discontinuities in the Membranes of Vesicles Using ImageJ. J Fluoresc 30, 735–740 (2020). https://doi.org/10.1007/s10895-020-02560-9

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Keywords

  • Edge detection
  • Membranes
  • Submicron discontinuities
  • Submicron pores
  • ImageJ