Sodium gradient-dependent phosphate transport in placental brush border membrane vesicles.

We recently described a sodium gradient-dependent transport of phosphate through the brush border membrane vesicles from human placenta. In order to characterize this transport carrier further, we studied the influence of temperature and membrane potential on the transport of this electrolyte, the stoichiometry of the sodium-phosphate interaction, and the interrelationship between phosphate uptake and other sodium-dependent systems. Temperature influenced phosphate uptake by changing the maximal velocity and the affinity of the carrier for the substrate. The Arrhenius plot for uptake velocity exhibited an abrupt breakpoint at 28.6 degrees C, suggesting that membrane fluidity is a factor in phosphate uptake. Increasing the sodium concentration in the incubation medium augmented the phosphate uptake according to a sigmoid curve, and the Hill plot analysis of these data indicates that at least two sodium ions are transported with each phosphate radical. The effect of membrane potential on phosphate uptake was studied by inducing potassium diffusion with valinomycin and by using various sodium salts with different anion conductance in the incubation medium. In both series of experiments, the inside-negative potential significantly enhanced phosphate uptake. We concluded that the phosphate-sodium cotransport is an electrogenic process, a conclusion which is compatible with the observation that at least two sodium ions accompany each phosphate radical. Glycine, alanine and proline all inhibited phosphate uptake according to an uncompetitive type of inhibition. In contrast, the addition of glucose to the incubation medium had no effect.