Interaction of albumins from different species with phospholipid liposomes. Multiple binding sites system

The interactions of three serum albumin species (rat, human, and bovine) with liposomes containing dimyristoylphosphatidylcholine, distearoylphosphatidylcholine or mixtures of both under different membrane fluidity conditions have been investigated using isothermal titration calorimetry and steady-state fluorescence anisotropy. Calorimetric titration studies of the binding of liposomes to the albumin species indicate in all cases exothermic processes with multiple sites of binding in the albumin molecules. Distinct saturation of the protein-lipid binding processes was observed at low or high molar lipid/protein ratio depending on the particular system. The thermodynamic parameters, including the association enthalpy and entropy, and the optimal values for the binding constants were thoroughly varied as a function of the number of identical binding sites, defining the best value of the parameter. Our experimental results, obtained using complementary biophysical techniques, provide experimental evidence for a significant difference in the association of the three protein species to phospholipid membranes. These observations also suggest a close relation between the binding parameters of the protein/lipid association and the lipid state of the phospholipid membranes.