Biotin uptake mechanisms in brush-border and basolateral membrane vesicles isolated from rabbit kidney cortex

1986 ◽  
Vol 856 (3) ◽  
pp. 471-481 ◽  
Author(s):  
René-Alexandre Podevin ◽  
Bernadette Barbarat
1988 ◽  
Vol 254 (5) ◽  
pp. F711-F718 ◽  
Author(s):  
P. T. Cheung ◽  
M. R. Hammerman

To define the mechanism by which glucose is transported across the basolateral membrane of the renal proximal tubular cell, we measured D-[14C]glucose uptake in basolateral membrane vesicles from rabbit kidney. Na+-dependent D-glucose transport, demonstrable in brush-border vesicles, could not be demonstrated in basolateral membrane vesicles. In the absence of Na+, the uptake of D-[14C]glucose in basolateral vesicles was more rapid than that of L-[3H]glucose over a concentration range of 1-50 mM. Subtraction of the latter from the former uptakes revealed a saturable process with apparent Km of 9.9 mM and Vmax of 0.80 nmol.mg protein-1.s-1. To characterize the transport component of D-glucose uptake in basolateral vesicles, we measured trans stimulation of 2 mM D-[14C]glucose entry in the absence of Na+. Trans stimulation could be effected by preloading basolateral vesicles with D-glucose, 2-deoxy-D-glucose, or 3-O-methyl-D-glucose, but not with L-glucose or alpha-methyl-D-glucoside. Trans-stimulated D-[14C]glucose uptake was inhibited by 0.1 mM phloretin or cytochalasin B but not phlorizin. In contrast, Na+-dependent D-[14C]glucose transport in brush-border vesicles was inhibited by phlorizin but not phloretin or cytochalasin B. Our findings are consistent with the presence of a Na+-independent D-glucose transporter in the proximal tubular basolateral membrane with characteristics similar to those of transporters present in nonepithelial cells.


1980 ◽  
Vol 238 (6) ◽  
pp. F452-F460 ◽  
Author(s):  
S. Grinstein ◽  
R. J. Turner ◽  
M. Silverman ◽  
A. Rothstein

The efflux of inorganic anions from purified brush border and basolateral membrane vesicles from dog kidney cortex was measured under equilibrium exchange conditions. Marked differences in temperature sensitivity and effects of inhibitors were found between the Cl and SO4 transport pathways and between the two types of membranes. SO4 transport in both brush border and basolateral membranes was markedly reduced by cooling, but significant inhibition by 4,4'–diisothiocyano-2,2'–disulfonic stilbene (DIDS) was only observed in basolateral vesicles. In contrast, Cl efflux from both types of vesicles was neither substantially inhibited by DIDS nor by lowering the temperature to 0 degrees C. Phosphate efflux from basolateral membrane vesicles was found to be only partially sensitive to DIDS. Attempts to label the stilbene-sensitive SO4 pathway in basolateral vesicles using [3H2]DIDS as a marker were unsuccessful due to the nonspecific labeling of many membrane components. The asymmetry in inorganic anion transport behavior exhibited by brush border and basolateral membrane vesicles from dog renal proximal tubule was also observed in equivalent vesicles prepared from rat small intestine.


1983 ◽  
Vol 244 (6) ◽  
pp. F686-F695 ◽  
Author(s):  
K. E. Jorgensen ◽  
U. Kragh-Hansen ◽  
H. Roigaard-Petersen ◽  
M. I. Sheikh

The mechanisms of tubular transport of citrate in renal basolateral and luminal membrane vesicles were studied under various experimental conditions. Both membrane preparations take up citrate by a Na+-dependent transport system, although with different characteristics. The uptake of citrate by basolateral membrane vesicles was insensitive to changes in membrane potential, which is indicative of electroneutral transport of the anion. The Na+-dependent uptake of citrate by luminal membrane vesicles was influenced by the presence of Na+salt anions of different permeabilities in the order: chloride greater than sulfate greater than gluconate. Furthermore, addition of citrate to membrane vesicle-potential-sensitive dye suspensions resulted in optical changes of the dye, indicative of electrogenic transfer of this compound. The apparent affinity of the citrate transport system located in luminal membrane vesicles, in contrast to basolateral membrane vesicles, was sensitive to changes in medium pH and was higher than that of basolateral membrane vesicles in the pH range studied. On the basis of these results a model for the transport of citrate by rabbit kidney proximal tubule is proposed.


1982 ◽  
Vol 208 (2) ◽  
pp. 377-382 ◽  
Author(s):  
M I Sheikh ◽  
U Kragh-Hansen ◽  
K E Jørgensen ◽  
H Røigaard-Petersen

A procedure for isolation and separation of purified luminal-membrane and basolateral-membrane vesicles from adult and newborn rabbit renal cortex by using Ca2+/Mg2+ precipitation, differential centrifugation and a self-orienting Percoll-gradient centrifugation is described. The purity of the membrane-vesicle suspensions was examined by electron microscopy and by measuring the activity of several marker enzymes. The activity of Na+ + K+-stimulated ATPase in the fraction mainly containing adult rabbit basolateral-membrane vesicles was enriched 16-fold, and the activity of alkaline phosphatase in the fraction mainly containing luminal-membrane vesicles was increased 13-fold, compared with the homogenate. Similar results were obtained with kidneys from newborn rabbits. Uptake studies, with a rapid filtration technique and the spectrophotometric method described in an accompanying paper [Kragh-Hansen, Jørgensen & Sheikh (1982) Biochem. J. 208, 359-368], showed that both adult and newborn rabbit luminal-membrane vesicles, in contrast with the basolateral-membrane preparations, possess an Na+-dependent electrogenic transport system for L-proline. Adult rabbit luminal-membrane vesicles take up citrate and L-malate by Na+-dependent electrogenic processes, whereas adult rabbit basolateral membrane vesicles do not exhibit electrogenic uptake of citrate. By contrast, these vesicles show Na+-dependent electrogenic uptake of L-malate.


1982 ◽  
Vol 208 (2) ◽  
pp. 359-368 ◽  
Author(s):  
Ulrich Kragh-Hansen ◽  
Karl Evald Jørgensen ◽  
M. Iqbal Sheikh

Renal transport of four different categories of organic solutes, namely sugars, neutral amino acids, monocarboxylic acids and dicarboxylic acids, was studied by using the potential-sensitive dye 3,3′-diethyloxadicarbocyanine iodide in purified luminal-membrane and basolateral-membrane vesicles isolated from rabbit kidney cortex. Valinomycin-induced K+ diffusion potentials resulted in concomitant changes in dye–membrane-vesicle absorption spectra. Linear relationships were obtained between these changes and depolarization and hyperpolarization of the vesicles. Addition of d-glucose, l-phenylalanine, succinate or l-lactate to luminal-membrane vesicles, in the presence of an extravesicular>intravesicular Na+ gradient, resulted in rapid transient depolarization. With basolateral-membrane vesicles no electrogenic transport of d-glucose or l-phenylalanine was observed. Spectrophotometric competition studies revealed that d-galactose is electrogenically taken up by the same transport system as that for d-glucose, whereas l-phenylalanine, succinate and l-lactate are transported by different systems in luminal-membrane vesicles. The absorbance changes associated with simultaneous addition of d-glucose and l-phenylalanine were additive. The uptake of these solutes was influenced by the presence of Na+-salt anions of different permeabilities in the order: Cl−>SO42−>gluconate. Addition of valinomycin to K+-loaded vesicles enhanced uptake of d-glucose and l-phenylalanine in the presence of an extravesicular>intravesicular Na+ gradient. Gramicidin or valinomycin plus nigericin diminished/abolished electrogenic solute uptake by Na+- or Na++K+-loaded vesicles respectively. These results strongly support the presence of Na+-dependent renal electrogenic transport of d-glucose, l-phenylalanine, succinate and l-lactate in luminal-membrane vesicles.


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