Interaction of phlorizin and sodium with the renal brush-border membrane D-glucose transporter: stoichiometry and order of binding

Abstract

The order and stoichiometry of the binding of phlorizin and sodium to the renal brush-border membrane D-glucose transporter are studied. The experimental results are consistent with a random-binding scheme in which the ratio of phlorizin- to sodium-binding sites is one-to-one. When the kinetics of phlorizin binding are measured as a function of increasing sodium concentration no significant variation is found in the apparent number of binding sites; however, the apparent binding constant for phlorizin decreases rapidly from approximately 16 microM at [Na] = 0 to 0.1 microM at [Na] = 100 mM and approaches 0.05 microM as [Na] leads to infinity. The experimental data are fit to a random carrier-type model of the coupled transport of sodium and D-glucose. A complete parameterization of the phlorizin binding properties of this model under sodium equilibrium conditions is given.