Electrogenic properties of the sodium-alanine cotransporter in pancreatic acinar cells: II. Comparison with transport models.

In this paper, the results of the preceding electrophysiological study of sodium-alanine cotransport in pancreatic acinar cells are compared with kinetic models. Two different types of transport mechanisms are considered. In the "simultaneous" mechanism the cotransporter C forms a ternary complex NCS with Na+ and the substrate S; coupled transport of Na+ and S involves a conformational transition between states NC'S and NC"S with inward- and outward-facing binding sites. In the "consecutive" (or "ping-pong") mechanism, formation of a ternary complex is not required; coupled transport occurs by an alternating sequence of association-dissociation steps and conformational transitions. It is shown that the experimentally observed alanine- and sodium-concentration dependence of transport rates is consistent with the predictions of the "simultaneous" model, but incompatible with the "consecutive" mechanism. Assuming that the association-dissociation reactions are not rate-limiting, a number of kinetic parameters of the "simultaneous" model can be estimated from the experimental results. The equilibrium dissociation constants of Na+ and alanine at the extracellular side are determined to be K"N less than or equal to 64 mM and K"S less than or equal to 18 mM. Furthermore, the ratio K"N/KS"N of the dissociation constants of Na+ from the binary (NC) and the ternary complex (NCS) at the extracellular side is estimated to be less than or equal to 6. This indicates that the binding sequence of Na+ and S to the transporter is not ordered. The current-voltage behavior of the transporter is analyzed in terms of charge translocations associated with the single-reaction steps. The observed voltage-dependence of the half-saturation concentration of sodium is consistent with the assumption that a Na+ ion that migrates from the extracellular medium to the binding site has to traverse part of the transmembrane voltage.