title: Lipid peroxidation and water penetration in lipid bilayers: a W-band EPR study.
abstract: Lipid peroxidation plays a key role in the alteration of cell membrane's properties. Here we used as model systems multilamellar vesicles (MLVs) made of the first two products in the oxidative cascade of linoleoyl lecithin, namely 1-palmitoyl-2-(13-hydroperoxy-9,11-octadecanedienoyl)-lecithin (HpPLPC) and 1-palmitoyl-2-(13-hydroxy-9,11-octadecanedienoyl)-lecithin (OHPLPC), exhibiting a hydroperoxide or a hydroxy group at position 13, respectively. The two oxidized lipids were used either pure or in a 1:1 molar ratio mixture with untreated 1-palmitoyl-2-linoleoyl-lecithin (PLPC). The model membranes were doped with spin-labeled lipids to study bilayer alterations by electron paramagnetic resonance (EPR) spectroscopy. Two different spin-labeled lipids were used, bearing the doxyl ring at position (n) 5 or 16: γ-palmitoyl-β-(n-doxylstearoyl)-lecithin (n-DSPPC) and n-doxylstearic acid (n-DSA). Small changes in the acyl chain order in the sub-polar region and at the methyl-terminal induced by lipid peroxidation were detected by X-band EPR. Concomitantly, the polarity and proticity of the membrane bilayer in those regions were investigated at W band in frozen samples. Analysis of the g(xx) and A(zz) parameters revealed that OHPLPC, but mostly HpPLPC, induced a measurable increase in polarity and H-bonding propensity in the central region of the bilayer. Molecular dynamics simulation performed on 16-DSA in the PLPC-HpPLPC bilayer revealed that water molecules are statistically favored with respect to the hydroperoxide groups to interact with the nitroxide at the methyl-terminal, confirming that the H-bonds experimentally observed are due to increased water penetration in the bilayer. The EPR and MD data on model membranes demonstrate that cell membrane damage by oxidative stress cause alteration of water penetration in the bilayer.
authors.affiliation: Centro di Studio sui Mitocondri e Metabolismo Energetico (CNR) c/o Dipartimento di Biochimica e Biologia Molecolare, Università di Bari, V. Orabona 4, 70126 Bari, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Conte
authors.forename: Francesco Maria
authors.initials: FM
authors.lastname: Megli
authors.forename: Himanshu
authors.initials: H
authors.lastname: Khandelia
authors.forename: Gunnar
authors.initials: G
authors.lastname: Jeschke
authors.forename: Enrica
authors.initials: E
authors.lastname: Bordignon
mesh_terms: D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D006860:Hydrogen Bonding; D054709:Lecithins / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D015227:Lipid Peroxidation; D008055:Lipids / Q000737:chemistry*; D008567:Membranes, Artificial; D008956:Models, Chemical; D056004:Molecular Dynamics Simulation; D010100:Oxygen / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D014867:Water / Q000737:chemistry*
chemical_list: D054709:Lecithins; D008051:Lipid Bilayers; D008055:Lipids; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D010743:Phospholipids; D014867:Water; C055402:1-palmitoyl-2-linoleoylphosphatidylcholine; D010100:Oxygen