Cationic amphiphiles and the solubilization of cholesterol crystallites in membrane bilayers.
Cationic amphiphiles used for transfection can be incorporated into biological membranes. By differential scanning calorimetry (DSC), cholesterol solubilization in phospholipid membranes, in the absence and presence of cationic amphiphiles, was determined. Two different systems were studied: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)+cholesterol (1:3, POPC:Chol, molar ratio) and 1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-l-serine] (POPS)+cholesterol (3:2, POPS:Chol, molar ratio), which contain cholesterol in crystallite form. For the zwitterionic lipid POPC, cationic amphiphiles were tested, up to 7 mol%, while for anionic POPS bilayers, which possibly incorporate more positive amphiphiles, the fractions used were higher, up to 23 mol%. 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and DOTAP in methyl sulfate salt form (DOTAPmss) were found to cause a small decrease on the enthalpy of the cholesterol transition of pure cholesterol aggregates, possibly indicating a slight increase on the cholesterol solubilization in POPC vesicles. With the anionic system POPS:Chol, the cationic amphiphiles dramatically change the cholesterol crystal thermal transition, indicating significant changes in the cholesterol aggregates. For structural studies, phospholipids spin labeled at the 5th or 16th carbon atoms were incorporated. In POPC, at the bilayer core, the cationic amphiphiles significantly increase the bilayer packing, decreasing the membrane polarity, with the cholesterol derivative 3 beta-[N-(N',N'-dimethylaminoethane)-carbamoyl]-cholesterol (DC-chol) displaying a stronger effect. In POPS and POPS:Chol, DC-chol was also found to considerably increase the bilayer packing. Hence, exogenous cationic amphiphiles used to deliver nucleic acids to cells can change the bilayer packing of biological membranes and alter the structure of cholesterol crystals, which are believed to be the precursors to atherosclerotic lesions.
Benatti
Carlos R
CR
Instituto de Física, Universidade de S. Paulo, CP 66318, CEP 05315-970, S. Paulo, SP, Brazil. crbenatti@gmail.com <crbenatti@gmail.com>
Lamy
M Teresa
MT
Epand
Richard M
RM
D002152:Calorimetry, Differential Scanning; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D012995:Solubility; D013696:Temperature; D013816:Thermodynamics
D005229:Fatty Acids, Monounsaturated; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D000644:Quaternary Ammonium Compounds; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; C050294:1-palmitoyl-2-oleoylglycero-3-phosphoserine; D002784:Cholesterol; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; C028694:1-palmitoyl-2-oleoylphosphatidylcholine