Phosphatidic acid distribution on the external surface of mixed vesicles

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Abstract
A new method was developed to detect the distribution of phosphatidic acid on the external surface of mixed phospholipid vesicles. Some positive dyes undergo large absorbance changes when the spatial separation between 2 or more dye molecules is smaller than a critical distance. When these dyes interact with mixed phospholipid vesicles, the absorbance changes may be utilized to calculate the amount of phosphatidic acid molecules which, on the external surface, occupy nearby positions not exceeding the critical dye distance, i.e., the amount of paired phosphatidic acid molecules. This amount was found to be higher than that calculated by statistical methods, indicating that phosphatidic acid molecules tend to be associated, in spite of the electrostatic repulsion between negative groups. The dependence of the amount of paired phosphatidic acid molecules on the pH, phosphatidylcholine:phosphatidic acid ratio and temperature was also analyzed.