Interactions of Dicarboxylic Porphyrins with Unilamellar Lipidic Vesicles: Drastic Effects of pH and Cholesterol on Kinetics

The effect of pH on the transfer of deuteroporphyrin from dimyristoylphosphatidylcholine (DMPC) unilamellar vesicles to human serum albumin is investigated using a stopped-flow with fluorescence detection. The kinetics of this process allows for the determination of the rate constants for the porphyrin exist from the outer vesicle layer to the bulk aqueous medium (koff), and for its movement from the inner to the outer vesicle layer (kto). Both koff and kto are found to strongly depend on the pH. The observed behavior can be described by classical titration curves and is most likely due to protonation equilibria involving the two carboxylic groups of the porphyrin. A pH increase accelerates the exist of the porphyrin. The reverse effect is observed for its movement through the bilayer. The presence of cholesterol in the DMPC bilayer also strongly affects the interactions of the porphyrin with the vesicles. The rate constant kto is dramatically reduced by increasing the cholesterol content. An irregularity is noted around 10-20 mol % cholesterol. The results are discussed in relation to the preferential uptake of porphyrins by tumors, a basis of photodynamic therapy, and to possible pH-mediated relocalization of porphyrins among subcellular structures.