Effect of the lipid phase transition on the kinetics of H+/OH- diffusion across phosphatidic acid bilayers

The kinetics of H+/OH- diffusion across dimyristoyl phosphatidic acid bilayer membranes was measured by following the absorbance of the pH-sensitive indicator Cresol red (o-cresolsulfonphthalein) entrapped in single lamellar vesicles after rapidly changing the external pH in a stopped-flow apparatus. The H+/OH- permeability coefficient was found to be in the 10(-5) to 10(-3) cm . s-1 range. The lipid phase transition has a strong influence on the permeation kinetics as the permeability coefficients in the liquid-crystalline phase are drastically higher. The permeability shows no maximum at the phase transition temperature as is the case for other ions, but displays a similar temperature dependence as water permeation. This is also reflected in the high activation energy of approx. 20 kcal/mol and supports the hypothesis (Nichols, J.W. and Deamer, D.W. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 2038-2042) of H+/OH- permeation via hydrogen bonded water molecules. A second slower kinetic phase is also observed, where the permeation is obviously controlled by counterion diffusion. The temperature dependence of this slow process displays the for ion diffusion characteristic maximum in the permeability at the phase-transition temperature.