RESUMO
Temperature-jump relaxation experiments on Na+ transport by (221)C10-cryptand (ionizable mobile carrier) and nonactin (neutral mobile carrier) were carried out in order to study the effects of cholesterol and the degree of acyl chain unsaturation, and their temperature-dependence on ion transport through thin lipid membranes. The experiments were performed on large, negatively charged unilamellar vesicles (LUV) prepared from mixtures of phosphatidylcholine (egg phosphatidylcholine, dioleoylphosphatidylcholine and dilinoleolylphosphatidylcholine), phosphatidic acid and cholesterol (mole fractions 0-0.43), at various temperatures and carrier concentrations. The apparent rate constants of Na+ translocation by (221)C10 and nonactin increased with the carrier concentration, the degree of acyl chain unsaturation and the temperature. The incorporation of cholesterol into the membranes significantly reduced the carrier concentration-, acyl chain unsaturation- and temperature-dependence of this parameter. The apparent energy required to activate the transport decreased significantly with increasing (221)C10 concentrations and remained constant with increasing those of nonactin at any given cholesterol molar fraction and degree of acyl chain unsaturation. It increased significantly with increasing the cholesterol molar fraction at any given carrier concentration to an extent depending on the degree of acyl chain unsaturation. Our interpretation of the action of cholesterol on these transport systems is based on the assumption that the adsorption plane of Na(+)-(221)C10 and Na(+)-nonactin complexes is likely to be located towards the aqueous and the hydrocarbon side of the dipole layer, respectively. The results are discussed in terms of the structural, physico-chemical and electrical characteristics of carriers and complexes, and of the interactions occurring between an ionizable or a neutral mobile carrier and the membrane.