Dicarboxylate transport in basolateral membrane vesicles prepared from rat kidney cortex was studied using 3H-methylsuccinate as a substrate. A sodium gradient (out greater than in) simulated methylsuccinate uptake and led to a transient overshoot. Lithium inhibited methylsuccinate uptake in the presence of sodium. The dependence of methylsuccinate uptake on sodium concentration indicated the interaction of more than one sodium ion with the transporter. Half-maximal stimulation was observed at 24 mmol/l sodium. Sodium-driven methylsuccinate uptake was electrogenic carrying a net positive charge. The basolateral dicarboxylate transport system exhibited an optimum at pH 7.0-7.5. In contrast, the sodium-dependent dicarboxylate transport system of brush border membranes depended much less on pH and had no optimum in the tested range. Cis-inhibition studies showed a preference of the system for dicarboxylates in the trans-configuration (fumarate) over cis-dicarboxylates (maleate). Citrate was accepted but oxalate and L-glutamate were not. DIDS exhibited a small inhibition. Among the monocarboxylates, gluconate and pyruvate inhibited methylsuccinate uptake whereas probenecid and p-aminohippurate (1 mmol/l) were without effect. The data indicate the presence of a sodium-dependent transport system in the basolateral membrane which accepts tricarboxylic acid cycle intermediates. This system is most likely not identical to the transport system responsible for organic anion secretion.