The Renal Sodium/Phosphate Symporters: Evidence for Different Functional Oligomeric States

Abstract

The oligomeric size of the rat renal sodium/phosphate symporters was estimated in brush-border membrane vesicles submitted to radiation inactivation. Altering the electrochemical conditions under which phosphate transport was measured resulted in different molecular size determinations. The radiation inactivation size (RIS) obtained from the radiation-induced loss of transport activity measured in the presence of a sodium gradient was 200 kDa. Under sodium equilibrium conditions, in the presence of a phosphate gradient as the only driving force, transport fell to 13% of the activity measured in the presence of a sodium gradient, and the RIS was 62 kDa. Addition of an outwardly-directed proton gradient increased the transport activity to 29% of that measured in the presence of a sodium gradient. The RIS measured under these conditions was 124 kDa. Under all conditions tested, phosphate uptake by irradiated vesicles was significantly reduced but remained linear during the first 5 s of incubation. The radiation-induced loss of transport activity was thus attributable to a direct inactivation of the transporter rather than to a decrease in the physical integrity of the vesicles. These results are consistent with a tetrameric structure composed of subunits of about 62 kDa and suggest that phosphate transport involves both monomers and tetramers.