Temperature-dependent relationship between K+ influx, Mg2+-ATPase activity, transmembrane potential and membrane...

The temperature-dependent relationship between K+ active influx, Mg2+-ATPase activity, transmembrane potential (delta psi) and the membrane lipid composition has been investigated in mycoplasma PG3. Native organisms were grown in a medium containing 10 microgram/ml cholesterol and either oleic plus palmitic (chol (+), O + P) or elaidic (chol (+), E) acids. Adapted cells were grown in a medium free of exogenous cholesterol and supplemented with elaidic acid (chol (-), E). Arrhenius plots of 42K+ active influx gave a linear relationship for (chol (+), O + P) cells (EA = -9 kcal). On the other hand, when oleic plus palmitic acids are replaced by elaidic acid, an upward discontinuity appears between 28 and 30 degrees C, which is associated with a large increase in the apparent activation energy of the process (t > 30 degrees C, EA = -24 kcal; t < 30 degrees C, EA = -40 kcal). Finally, a biphasic response with a break at approx. 23 degrees C (EA = -7 kcal, t > 23 degrees C; EA = -44 kcal, t < 23 degrees C) is observed for (chol(-), E) organisms. From the lack of correspondence between these effects on the K+ influx and the temperature dependence of both the Mg2+-ATPase activity and delta psi, it is suggested that changes in the membrane lipid composition affect the K+ transport at the level of the K+ carrier itself. Differential scanning calorimetry, steady-state fluorescence polarization of diphenylhexatriene and freeze-fracture electron microscopy experiments further suggest that the effect is largely due to modifications of the membrane microviscosity and that the K+ carrier is associated with the most fluid lipid species present in the membrane.