title: Isothermal titration calorimetric analysis of the interaction between cationic lipids and plasmid DNA.
abstract: The effects of buffer and ionic strength upon the enthalpy of binding between plasmid DNA and a variety of cationic lipids used to enhance cellular transfection were studied using isothermal titration calorimetry at 25.0 degrees C and pH 7.4. The cationic lipids DOTAP (1,2-dioleoyl-3-trimethyl ammonium propane), DDAB (dimethyl dioctadecyl ammonium bromide), DOTAP:cholesterol (1:1), and DDAB:cholesterol (1:1) bound endothermally to plasmid DNA with a negligible proton exchange with buffer. In contrast, DOTAP: DOPE (L-alpha-dioleoyl phosphatidyl ethanolamine) (1:1) and DDAB:DOPE (1:1) liposomes displayed a negative enthalpy and a significant uptake of protons upon binding to plasmid DNA at neutral pH. These findings are most easily explained by a change in the apparent pKa of the amino group of DOPE upon binding. Complexes formed by reverse addition methods (DNA into lipid) produced different thermograms, sizes, zeta potentials, and aggregation behavior, suggesting that structurally different complexes were formed in each titration direction. Titrations performed in both directions in the presence of increasing ionic strength revealed a progressive decrease in the heat of binding and an increase in the lipid to DNA charge ratio at which aggregation occurred. The unfavorable binding enthalpy for the cationic lipids alone and with cholesterol implies an entropy-driven interaction, while the negative enthalpies observed with DOPE-containing lipid mixtures suggest an additional contribution from changes in protonation of DOPE.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence 66047, USA.
authors.forename: B A
authors.initials: BA
authors.lastname: Lobo
authors.forename: A
authors.initials: A
authors.lastname: Davis
authors.forename: G
authors.initials: G
authors.lastname: Koe
authors.forename: J G
authors.initials: JG
authors.lastname: Smith
authors.forename: C R
authors.initials: CR
authors.lastname: Middaugh
mesh_terms: D002151:Calorimetry / Q000379:methods*; D002412:Cations; D004247:DNA / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D020404:Glycerophospholipids / Q000737:chemistry; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007700:Kinetics; D008027:Light; D050356:Lipid Metabolism; D008081:Liposomes / Q000378:metabolism; D010714:Phosphatidylethanolamines; D010957:Plasmids / Q000378:metabolism*; D011485:Protein Binding; D011522:Protons; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012542:Scattering, Radiation; D012965:Sodium Chloride / Q000494:pharmacology; D013696:Temperature; D013816:Thermodynamics; D013997:Time Factors
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D002412:Cations; D005229:Fatty Acids, Monounsaturated; D020404:Glycerophospholipids; D007477:Ions; D008081:Liposomes; D010714:Phosphatidylethanolamines; D011522:Protons; D000644:Quaternary Ammonium Compounds; C046112:didodecyldimethylammonium; D012965:Sodium Chloride; D004247:DNA; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane