The Journal of Membrane Biology

, Volume 251, Issue 3, pp 475–489 | Cite as

Effect of 5-trans Isomer of Arachidonic Acid on Model Liposomal Membranes Studied by a Combined Simulation and Experimental Approach

  • Ioanna Tremi
  • Dimitrios Anagnostopoulos
  • Ellas Spyratou
  • Paraskevi Gkeka
  • Alexandros G. GeorgakilasEmail author
  • Chryssostomos ChatgilialogluEmail author
  • Zoe CourniaEmail author
Part of the following topical collections:
  1. Lipid Membranes and Reactions at Lipid Interfaces: Theory, experiments, and applications


Unsaturated fatty acids are found in humans predominantly in the cis configuration. Fatty acids in the trans configuration are primarily the result of human processing (trans fats), but can also be formed endogenously by radical stress. The cis–trans isomerization of fatty acids by free radicals could be connected to several pathologies. Trans fats have been linked to an increased risk of coronary artery disease; however, the reasons for the resulting pathogenesis remain unclear. Here, we investigate the effect of a mono-trans isomer of arachidonic acid (C20:4-5trans, 8cis, 11cis, 14cis) produced by free radicals in physiological concentration on a model erythrocyte membrane using a combined experimental and theoretical approach. Molecular Dynamics (MD) simulations of two model lipid bilayers containing arachidonic acid and its 5-trans isomer in 3 mol% were carried out for this purpose. The 5-trans isomer formation in the phospholipids was catalyzed by HOCH2CH2S· radicals, generated from the corresponding thiol by γ-irradiation, in multilamellar vesicles of SAPC. Large unilamellar vesicles were made by the extrusion method (LUVET) as a biomimetic model for cistrans isomerization. Atomic Force Microscopy and Dynamic Light Scattering were used to measure the average size, morphology, and the z-potential of the liposomes. Both results from MD simulations and experiments are in agreement and indicate that the two model membranes display different physicochemical properties in that the bilayers containing the trans fatty acids were more ordered and more rigid than those containing solely the cis arachidonic acid. Correspondingly, the average size of the liposomes containing trans isomers was smaller than the ones without.


Trans fatty acids Arachidonic acid Liposomes MD simulations 



We acknowledge computational time granted from the Greek Research & Technology Network (GRNET) in the National HPC facility – ARIS under project ID pr001026-MAG-NANO-MEM. We also acknowledge Dr. Eleni Efthimiadou for providing us the Zetasizer for the Dynamic Light Scattering experiments and Giorgia Giacometti for her ancillary help in the liposome preparation. ZC was co-funded by the European Commission under the H2020 Research Infrastructures Contract No. 675121 (Project VI-SEEM). All data is available to download at:

Compliance with Ethical Standards

Conflict of interest

All authors declare that they have no conflict of interest.

Research Involving Human and Animal Participants

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

232_2018_29_MOESM1_ESM.pdf (2 mb)
Supplementary material 1 (PDF 2047 KB)


  1. Aksimentiev A, Schulten K (2005) Imaging alpha-hemolysin with molecular dynamics: ionic conductance, osmotic permeability, and the electrostatic potential map. Biophys J 88(6):3745–3761CrossRefPubMedPubMedCentralGoogle Scholar
  2. Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P (2002) Molecular biology of the cell. Garland Science, New YorkGoogle Scholar
  3. Angelikopoulos P, Sarkisov L, Cournia Z, Gkeka P (2017) Self-assembly of anionic ligand-coated nanoparticles in lipid membranes. Nanoscale 9(3):1040–1048CrossRefPubMedGoogle Scholar
  4. Bar-Ziv R, Moses E, Nelson P (1998) Dynamic excitations in membranes induced by optical tweezers. Biophys J 75(1):294–320CrossRefPubMedPubMedCentralGoogle Scholar
  5. Barber CB, Dobkin DP, Huhdanpaa H (1996) The quickhull algorithm for convex hulls. ACM Trans Math Softw 22(4):469–483CrossRefGoogle Scholar
  6. Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O (2012) Oxidative stress antioxidant defense. World Allergy Org J 5(1):9–19CrossRefGoogle Scholar
  7. Cappella B, Dietler G (1999) Force-distance curves by atomic force microscopy. Surf Sci Rep 34(1):1–104CrossRefGoogle Scholar
  8. Chaban V (2014) Computationally efficient prediction of area per lipid. Chem Phys Lett 616:25–29CrossRefGoogle Scholar
  9. Chatgilialoglu C, Ferreri C (2005) Trans lipids: the free radical path. Acc Chem Res 38(6):441–448CrossRefPubMedGoogle Scholar
  10. Chatgilialoglu C, Ferreri C (2010) Biomimetic chemistry: radical reactions in vesicle suspensions. In: mukherjee A Biomimetics learning from nature, InTech, RijekaGoogle Scholar
  11. Chatgilialoglu C, Ferreri C, Ballestri M, Mulazzani QG, Landi L (2000) cis–trans isomerization of monounsaturated fatty acid residues in phospholipids by thiyl radicals. J Am Chem Soc 122(19):4593–4601CrossRefGoogle Scholar
  12. Chatgilialoglu C, Ferreri C, Lykakis IN, Mihaljević B (2014a) Biomimetic thiyl radical chemistry by γ-irradiation of micelles and vesicles containing unsaturated fatty acids. Isr J Chem 54(3):242–247CrossRefGoogle Scholar
  13. Chatgilialoglu C, Ferreri C, Melchiorre M, Sansone A, Torreggiani A (2014b) Lipid geometrical isomerism: from chemistry to biology and diagnostics Chem Rev 114(1):255–284CrossRefPubMedGoogle Scholar
  14. Cort A, Ozben T, Melchiorre M, Chatgilialoglu C, Ferreri C, Sansone A (2016) Effects of bleomycin and antioxidants on the fatty acid profile of testicular cancer cell membranes. Biochim et Biophys Acta 1858(2):434–441CrossRefGoogle Scholar
  15. Cournia Z, Allen TW, Andricioaei I, Antonny B, Baum D, Brannigan G, Buchete N-V, Deckman JT, Delemotte L, del Val C, Friedman R, Gkeka P, Hege H-C, Hénin J, Kasimova MA, Kolocouris A, Klein ML, Khalid S, Lemieux MJ, Lindow N, Roy M, Selent J, Tarek M, Tofoleanu F, Vanni S, Urban S, Wales DJ, Smith JC, Bondar A-N (2015) Membrane protein structure, function, and dynamics: a perspective from experiments and theory. J Membr Biol 248(4):611–640CrossRefPubMedPubMedCentralGoogle Scholar
  16. Cournia Z, Ullmann GM, Smith JC (2007) Differential effects of cholesterol, ergosterol and lanosterol on a dipalmitoyl phosphatidylcholine membrane: a molecular dynamics simulation study. J Phys Chem B 111(7):1786–1801CrossRefPubMedGoogle Scholar
  17. Egwim PO, Sgoutas DS (1971) Occurrence of eicosadienoic acids in liver lipids of rats fed partially hydrogenated soybean fat. J Nutr 101(3):307–314CrossRefPubMedGoogle Scholar
  18. Emken EA, Adlof RO, Rohwedder WK, Gulley RM (1983) Incorporation of deuterium-labeled trans- and cis-13-octadecenoic acids in human plasma lipids. J Lipid Res 24(1):34–46PubMedGoogle Scholar
  19. Feller SE, Zhang Y, Pastor RW, Brooks BR (1995) Constant pressure molecular dynamics simulation: the langevin piston method. J Chem Phys 103(11):4613–4621CrossRefGoogle Scholar
  20. Ferreri C, Chatgilialoglu C (2012) Role of fatty acid-based functional lipidomics in the development of molecular diagnostic tools. Expert Rev Mol Diagn 12(7):767–780CrossRefPubMedGoogle Scholar
  21. Ferreri C, Costantino C, Perrotta L, Landi L, Mulazzani QG, Chatgilialoglu C (2001) Cis-trans isomerization of polyunsaturated fatty acid residues in phospholipids catalyzed by thiyl radicals. J Am Chem Soc 123(19):4459–4468CrossRefPubMedGoogle Scholar
  22. Ferreri C, Mennella MR, Formisano C, Landi L, Chatgilialoglu C (2002) Arachidonate geometrical isomers generated by thiyl radicals: the relationship with trans lipids detected in biological samples. Free Radic Biol Med 33(11):1516–1526CrossRefPubMedGoogle Scholar
  23. Ferreri C, Kratzsch S, Landi L, Brede O (2005) Thiyl radicals in biosystems: effects on lipid structures and metabolisms. Cell Mol Life Sci CMLS 62(7):834–847CrossRefPubMedGoogle Scholar
  24. Ferreri C, Samadi A, Sassatelli F, Landi L, Chatgilialoglu C (2004) Regioselective cis-trans isomerization of arachidonic double bonds by thiyl radicals: the influence of phospholipid supramolecular organization. J Am Chem Soc 126(4):1063–1072CrossRefPubMedGoogle Scholar
  25. Gardner HW (1989) Oxygen radical chemistry of polyunsaturated fatty acids. Free Radic Biol Med 7(1):65–86CrossRefPubMedGoogle Scholar
  26. Giacometti G, Marini M, Papadopoulos K, Ferreri C, Chatgilialoglu C (2017). trans-double bond-containing liposomes as potential carriers for drug delivery. Molecules 22(12)Google Scholar
  27. Gorter E, Grendel F (1925) On bimolecular layers of lipoids on the chromocytes of the blood. J Exp Med 41(4):439–443CrossRefPubMedPubMedCentralGoogle Scholar
  28. Guixa-Gonzalez R, Rodriguez-Espigares I, Ramirez-Anguita JM, Carrio-Gaspar P, Martinez-Seara H, Giorgino T, Selent J (2014) MEMBPLUGIN: studying membrane complexity in VMD. Bioinformatics 30(10):1478–1480CrossRefPubMedGoogle Scholar
  29. Humphrey W, Dalke A, Schulten K (1996) VMD: visual molecular dynamics. J Mol Graph 14(1):33–38, 27–38CrossRefPubMedGoogle Scholar
  30. Hyvönen M, Kovanen P (2005). Molecular dynamics simulations of unsaturated lipid bilayers: effects of varying the numbers of double bonds. Eur Biophys J 34(4):294–305CrossRefGoogle Scholar
  31. Ibarguren M, López DJ, Escribá PV (2014) The effect of natural and synthetic fatty acids on membrane structure, microdomain organization, cellular functions and human health. Biochim et Biophys Acta 1838(6):1518–1528CrossRefGoogle Scholar
  32. Janosi L, Gorfe AA (2010) Simulating POPC and POPC/POPG bilayers: conserved packing and altered surface reactivity. J Chem Theory Comput 6(10):3267–3273CrossRefPubMedGoogle Scholar
  33. Jo S, Kim T, Im W (2007) Automated builder and database of protein/membrane complexes for molecular dynamics simulations. PLoS ONE 2(9):e880CrossRefPubMedPubMedCentralGoogle Scholar
  34. Jorgensen WL, Chandrasekhar J, Madura JD, Impey RW, Klein ML (1983) Comparison of simple potential functions for simulating liquid water. J Chem Phys 79(2):926–935CrossRefGoogle Scholar
  35. Judd JT, Clevidence BA (1993) Trans fatty acids in human nutrition. Acta Cardiol 48(5):459–462PubMedGoogle Scholar
  36. Klauda JB, Venable RM, Freites JA, O’Connor JW, Tobias DJ, Mondragon-Ramirez C, Vorobyov I, MacKerell AD Jr, Pastor RW (2010) Update of the CHARMM all-atom additive force field for lipids: validation on six lipid types. J Phys Chem B 114(23):7830–7843CrossRefPubMedPubMedCentralGoogle Scholar
  37. Kučerka N, Nieh M-P, Katsaras J (2011) Fluid phase lipid areas and bilayer thicknesses of commonly used phosphatidylcholines as a function of temperature. Biochim et Biophys Acta 1808(11):2761–2771CrossRefGoogle Scholar
  38. Kučerka N, Tristram-Nagle S, Nagle JF (2006) Structure of fully hydrated fluid phase lipid bilayers with monounsaturated chains. J Membr Biol 208(3):193–202CrossRefGoogle Scholar
  39. Kulig W, Pasenkiewicz-Gierula M, Róg T (2016) Cis and trans unsaturated phosphatidylcholine bilayers: a molecular dynamics simulation study. Chem Phys Lipid 195:12–20CrossRefGoogle Scholar
  40. Lauritzen L, Hansen HS, Jorgensen MH, Michaelsen KF (2001) The essentiality of long chain n-3 fatty acids in relation to development and function of the brain and retina. Prog Lipid Res 40(1–2):1–94CrossRefPubMedGoogle Scholar
  41. Liang X, Mao G, Ng KY (2004) Mechanical properties and stability measurement of cholesterol-containing liposome on mica by atomic force microscopy. J Colloid Interface Sci 278(1):53–62CrossRefPubMedGoogle Scholar
  42. Lichtenstein AH (2000) Dietary trans fatty acid. J Cardiopulm Rehabil 20(3):143–146CrossRefPubMedGoogle Scholar
  43. Lodish HF (2013) Molecular Cell Biology. xxxiii 1154:1158Google Scholar
  44. Martyna GJ, Tobias DJ, Klein ML (1994) Constant pressure molecular dynamics algorithms. J Chem Phys 101(5):4177–4189CrossRefGoogle Scholar
  45. Matsuzaki K, Murase O, Sugishita Ki, Yoneyama S, Akada Ky, Ueha M, Nakamura A, Kobayashi S (2000) Optical characterization of liposomes by right angle light scattering and turbidity measurement. Biochim et Biophys Acta 1467(1):219–226CrossRefGoogle Scholar
  46. Melchiorre M, Torreggiani A, Chatgilialoglu C, Ferreri C (2011) Lipid markers of “geometrical” radical stress: synthesis of monotrans cholesteryl ester isomers and detection in human plasma. J Am Chem Soc 133(38):15184–15190CrossRefPubMedGoogle Scholar
  47. Mendonça MA, Araújo WMC, Borgo LA, E d. R Alencar (2017) Lipid profile of different infant formulas for infants. PLoS ONE 12(6):e0177812CrossRefPubMedPubMedCentralGoogle Scholar
  48. Murakami Y, Tsuyama M, Kobayashi Y, Kodama H, Iba K (2000) Trienoic fatty acids plant tolerance of high temperature. Science 287(5452):476–479CrossRefPubMedGoogle Scholar
  49. Murzyn K, Rog T, Jezierski G, Takaoka Y, Pasenkiewicz-Gierula M (2001) Effects of phospholipid unsaturation on the membrane/water interface: a molecular simulation study. Biophys J 81(1):170–183CrossRefPubMedPubMedCentralGoogle Scholar
  50. Neil W, Blackstone Rb (2007) The cell: a molecular approach. Fourth Edition. By Geoffrey M Cooper and Robert E Hausman. Quart Rev Biol 82(1):44–44Google Scholar
  51. New RRC (1990) Liposomes: a practical approach. IRL Press, OxfordGoogle Scholar
  52. Norris DO, Carr JA (2013) Chapter 3 - Synthesis, Metabolism, and Actions of Bioregulators. Vertebrate Endocrinology (Fifth Edition). Academic Press, San Diego, pp 41–91Google Scholar
  53. Petrache HI, Tu K, Nagle JF (1999) Analysis of simulated NMR order parameters for lipid bilayer structure determination. Biophys J 76(5):2479–2487CrossRefPubMedPubMedCentralGoogle Scholar
  54. Phillips JC, Braun R, Wang W, Gumbart J, Tajkhorshid E, Villa E, Chipot C, Skeel RD, Kale L, Schulten K (2005) Scalable molecular dynamics with NAMD. J Comput Chem 26(16):1781–1802CrossRefPubMedPubMedCentralGoogle Scholar
  55. Porasso RD, López Cascales JJ (2012) A criterion to identify the equilibration time in lipid bilayer simulations. Pap Phys 4(2):1–9Google Scholar
  56. Puca AA, Andrew P, Novelli V, Anselmi CV, Somalvico F, Cirillo NA, Chatgilialoglu C, Ferreri C (2007) Fatty acid profile of erythrocyte membranes as possible biomarker of longevity. Rejuvenation Res 11(1):63–72CrossRefGoogle Scholar
  57. Puca AA, Andrew P, Novelli V, Anselmi CV, Somalvico F, Cirillo NA, Chatgilialoglu C, Ferreri C (2008) Fatty acid profile of erythrocyte membranes as possible biomarker of longevity. Rejuvenation Res 11(1):63–72CrossRefPubMedGoogle Scholar
  58. 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(1):328–339CrossRefPubMedPubMedCentralGoogle Scholar
  59. Reddy AS, Warshaviak DT, Chachisvilis M (2012) Effect of membrane tension on the physical properties of DOPC lipid bilayer membrane. Biochim Biophys Acta 1818(9):2271–2281CrossRefPubMedPubMedCentralGoogle Scholar
  60. Ricciotti E, FitzGerald GA (2011) Prostaglandins inflammation. Arterioscler Thromb Vasc Biol 31(5):986–1000CrossRefPubMedPubMedCentralGoogle Scholar
  61. Roach C, Feller SE, Ward JA, Shaikh SR, Zerouga M, Stillwell W (2004) Comparison of cis and trans fatty acid containing phosphatidylcholines on membrane properties. Biochemistry 43(20):6344–6351CrossRefPubMedGoogle Scholar
  62. Ruozi B, Belletti D, Tombesi A, Tosi G, Bondioli L, Forni F, Vandelli MA (2011) AFM, ESEM, TEM, and CLSM in liposomal characterization: a comparative study. Int J Nanomed 6:557–563CrossRefGoogle Scholar
  63. Ruozi B, Tosi G, Leo E, Vandelli MA (2007) Application of atomic force microscopy to characterize liposomes as drug and gene carriers. Talanta 73(1):12–22CrossRefPubMedGoogle Scholar
  64. Samad A, Sultana Y, Aqil M (2007) Liposomal drug delivery systems: an update review. Curr Drug Deliv 4(4):297–305CrossRefPubMedGoogle Scholar
  65. Sansone A, Melchiorre M, Chatgilialoglu C, Ferreri C (2013) Hexadecenoic fatty acid isomers: a chemical biology approach for human plasma biomarker development. Chem Res Toxicol 26(11):1703–1709CrossRefPubMedGoogle Scholar
  66. Sansone A, Tolika E, Louka M, Sunda V, Deplano S, Melchiorre M, Anagnostopoulos D, Chatgilialoglu C, Formisano C, Di Micco R, Faraone Mennella MR, Ferreri C (2016) Hexadecenoic Fatty acid isomers in human blood lipids and their relevance for the interpretation of lipidomic profiles. PLOS ONE 11(4):e0152378CrossRefPubMedPubMedCentralGoogle Scholar
  67. Siegel G, Ermilov E, Pries AR, Winkler K, Schmidt A, Ringstad L, Malmsten M, Lindman B (2014). The significance of lipid peroxidation in cardiovascular disease. Colloids Surf A 442(Supplement C):173–180CrossRefGoogle Scholar
  68. Sneddon IN (1965) The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile. Int J Eng Sci 3(1):47–57CrossRefGoogle Scholar
  69. Soni SP, Ward JA, Sen SE, Feller SE, Wassall SR (2009) Effect of trans unsaturation on molecular organization in a phospholipid membrane. Biochemistry 48(46):11097–11107CrossRefPubMedGoogle Scholar
  70. Spyratou E, Mourelatou EA, Makropoulou M, Demetzos C (2009) Atomic force microscopy: a tool to study the structure, dynamics and stability of liposomal drug delivery systems. Expert Opin Drug Deliv 6(3):305–317CrossRefPubMedGoogle Scholar
  71. Stillwell W, Wassall SR (2003) Docosahexaenoic acid: membrane properties of a unique fatty acid. Chem Phys Lipid 126(1):1–27CrossRefGoogle Scholar
  72. van Meer G, Voelker DR, Feigenson GW (2008) Membrane lipids: where they are and how they behave. Nat Rev Mol Cell Biol 9(2):112–124CrossRefPubMedPubMedCentralGoogle Scholar
  73. Venable RM, Sodt AJ, Rogaski B, Rui H, Hatcher E, MacKerell AD, Pastor RW, Klauda B (2014) Charmm all-atom additive force field for sphingomyelin: elucidation of hydrogen bonding and of positive curvature. Biophys J 107(1):134–145CrossRefPubMedPubMedCentralGoogle Scholar
  74. Vermeer LS, de Groot BL, Réat V, Milon A, Czaplicki J (2007) Acyl chain order parameter profiles in phospholipid bilayers: computation from molecular dynamics simulations and comparison with 2H NMR experiments. Eur Biophys J 36(8):919–931CrossRefPubMedGoogle Scholar
  75. Wang Y, Gkeka P, Fuchs JE, Liedl KR, Cournia Z (2016) DPPC-cholesterol phase diagram using coarse-grained molecular dynamics simulations. Biochim et Biophys Acta 1858(11):2846–2857CrossRefGoogle Scholar
  76. Wolff RL, Entressangles B (1994) Steady-state fluorescence polarization study of structurally defined phospholipids from liver mitochondria of rats fed elaidic acid. Biochim Biophys Acta 1211(2):198–206CrossRefPubMedGoogle Scholar
  77. Wu EL, Cheng X, Jo S, Rui H, Song KC, Davila-Contreras EM, Qi Y, Lee J, Monje-Galvan V, Venable RM, Klauda JB, Im W (2014) CHARMM-GUI membrane Builder toward realistic biological membrane simulations. J Comput Chem 35(27):1997–2004CrossRefPubMedPubMedCentralGoogle Scholar
  78. Yarla NS, Bishayee A, Sethi G, Reddanna P, Kalle AM, Dhananjaya BL, Dowluru KSVG., Chintala R, Duddukuri GR (2016). Targeting arachidonic acid pathway by natural products for cancer prevention and therapy. Semin Cancer BiolGoogle Scholar
  79. Zambonin L, Ferreri C, Cabrini L, Prata C, Chatgilialoglu C, Landi L (2006) Occurrence of trans fatty acids in rats fed a trans-free diet: a free radical-mediated formation? Free Radic Biol Med 40(9):1549–1556CrossRefPubMedGoogle Scholar
  80. Zervou M, Cournia Z, Potamitis C, Patargias G, Durdagi S, Grdadolnik SG, Mavromoustakos T (2014) Insights into the molecular basis of action of the AT1 antagonist losartan using a combined NMR spectroscopy and computational approach. Biochim et Biophys Acta 1838(3):1031–1046CrossRefGoogle Scholar
  81. Zhu J, Sabharwal T, Guo L, Kalyanasundaram A, Wang G (2009) Gloss phenomena image analysis of atomic force microscopy in molecular cell biology. Scanning 31(2):49–58CrossRefPubMedPubMedCentralGoogle Scholar

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Authors and Affiliations

  1. 1.DNA Damage Laboratory, Department of Physics, School of Applied Mathematical and Physical SciencesNational Technical University of Athens (NTUA)AthensGreece
  2. 2.Biomedical Research Foundation Academy of AthensAthensGreece
  3. 3.Institute of Nanoscience and Nanotechnology (INN)N.C.S.R. DemokritosAthensGreece
  4. 4.ISOFConsiglio Nazionale delle RicercheBolognaItaly
  5. 5.GlaxoSmithKlineAthensGreece
  6. 6.Structure Design and InformaticsSanofiChilly-MazarinFrance

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