Nature and kinetic characteristics of L-DOPA uptake in rat renal proximal tubules

Abstract

The present study was performed with the aim to determine the kinetics and the caracteristics of cellular uptake of L-3,4-dihydroxyphenylalanine (L-DOPA) in rat renal proximal tubules. Incubation of renal tubules at 4°C in the presence of increasing concentrations of L-DOPA results in a linear and concentration-dependent accumulation of the substrate. In experiments carried out at 37°C, the accumulation of L-DOPA in renal tubules was found to be greater than that occurring at 4°C and showed a trend for saturation. The saturable component of L-DOPA uptake was derived from the total amount of L-DOPA accumulated in renal tubules at 37°C subtracted with the values obtained in experiments conducted at 4°C. The Vmax and Km values for the saturable component of L-DOPA uptake in renal tubules were, respectively, 241 ± 32 fmol µg protein(-1)min(-1) and 567 ± 63 µM. Cyanine 863 (5 and 10 µM) was found to decrease the tubular uptake of L-DOPA, whereas probenecid (50 µM) did not change the rate of uptake of L-DOPA into renal tubules. The Vmax and Km values for the saturable component of L-DOPA uptake in renal tubules incubated in the presence of 10 µM cyanine 863 were, respectively, 97 ± 11 fmol µg protein(-1)min(-1) and 160 ± 22 µM. It is suggested that the anionic L-DOPA may behave as an amphoteric substance, both hydroxyl groups in the aromatic ring determining the binding of the molecule to the organic cation transporter.