Sodium-sugar Coupling Stoichiometry in Chick Intestinal Cells

Sodium-dependent sugar transport systems involve the function of membrane components that couple the transmembrane flow of Na+ to the concomitant flow of certain sugar molecules. The coupling stoichiometry between Na+ and sugar fluxes via these systems must be measured under conditions in which the membrane potential does not change due to the induction of transport or during the interval of flux measurement. This can be accomplished by utilizing gradients of highly permeant ions (NO-3 and K+ plus valinomycin) to create diffusion potentials of sufficient magnitude that the sugar-induced Na+ flux does not introduce an appreciable change in the imposed potential. Under these conditions, the coupling stoichiometry for chicken intestinal cells proves to be 2 Na+:1 sugar as reported earlier for studies performed in the absence of a membrane potential. When control of the potential is not maintained, a coupling ratio of 1:1 is observed. The stoichiometry does not change as a function of Na+ concentration, which suggests that carrier forms with only one Na+ bound do not contribute to the carrier-mediated Na+ or sugar fluxes. When no potential is present, the stoichiometry is modified by the level of intracellular Na+ and sugar in a manner indicative of a transport mechanism in which Na+ must dissociate from the "loaded" carrier at the inward facing membrane surface before the sugar molecule dissociates.