Heterogeneity in the effects of membrane potentials on pantothenate and glucose uptakes by rabbit renal apical membranes.
1. Previous studies using renal brush-border membrane vesicles have established that both the pantothenate and the low Km (Michaelis-Menten constant), low Vmax (maximal rate) D-glucose systems have a stoichiometry of 2 Na+: 1 organic molecule. In this study, we compared the mechanisms by which the membrane potential energizes pantothenate and D-glucose uptakes by brush-border membrane vesicles isolated from the whole cortex of rabbit kidney. 2. In the absence of Na+, varying the membrane potential from +60 to -60 mV decreased pantothenate uptake, whereas D-glucose uptake was increased in a linear manner. These results suggested the existence of a conductive pathway for pantothenate in these membranes. They also suggested that the pantothenate free carrier is electroneutral, while the glucose free carrier is negatively charged. 3. In the presence of an inwardly directed Na+ gradient, varying the membrane potential from +60 to -60 mV increased Na(+)-dependent pantothenate influx linearly. In contrast, a shift from +60 to +40 mV in the membrane potential had no influence on Na(+)-dependent D-glucose influx, whereas influx was a linear function of the membrane potential from +40 to -60 mV, indicating that there is a threshold membrane potential required for membrane potential-dependent D-glucose movement to occur. 4. Kinetic studies revealed that the effect of membrane potential on pantothenate uptake is through changes in the Km, while Vmax was unchanged. On the other hand, the membrane potential exerted its effect on D-glucose transport solely on the Vmax. 5. Finally, binding studies revealed that membrane potential, both in the presence and absence of a Na+ gradient, elicited effects on phlorizin binding qualitatively similar to those observed for D-glucose transport. 6. Implications of these findings for tubular regulation of these electrogenic secondary active transport systems are discussed.
Barbarat
B
B
Laboratoire de Physiologie et Endocrinologie Cellulaire Rénale, Faculté de Médecine Broussais-Hotel Dieu, Paris, France.
Chambrey
R
R
Podevin
R A
RA
D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D005947:Glucose / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008564:Membrane Potentials / Q000502:physiology; D008871:Microvilli / Q000378:metabolism; D010205:Pantothenic Acid / Q000378:metabolism*; D010695:Phlorhizin / Q000378:metabolism; D011817:Rabbits; D012964:Sodium / Q000378:metabolism
D010205:Pantothenic Acid; D012964:Sodium; D010695:Phlorhizin; D005947:Glucose