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p-Aminohippurate/2-oxoglutarate exchange in bovine renal brush-border and basolateral membrane vesicles

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Abstract

The transport of the amphiphilic organic anion, P-aminohippurate (PAH), across the luminal (brush-border) and contraluminal (basolateral) membrane of renal proximal tubule cells was studied with membrane vesicles isolated from bovine kidney cortex. On the basis of the enrichment of specific activities of marker enzymes, leucine aminopeptidase and Na+/K+-ATPase, brush-border and basolateral membrane vesicles can be obtained from bovine kidneys in reasonably pure form. The uptake of [3H]PAH into both brush-border and basolateral membrane vesicles was trans-stimulated by intravesicular PAH and by 2-oxoglutarate. In the absence of Na+, [3H]PAH/2-oxoglutarate exchange was cis-inhibited by unlabelled 2-oxoglutarate in the medium. In the presence of an inward Na+ gradient, 10 μM 2-oxoglutarate, but no other Krebs cycle derivative, cis-stimulated [3H]PAH uptake, indicating that a Na3-coupled dicarboxylate transporter and PAH/2-oxoglutarate exchanger cooperate in both membranes to enhance [3H]PAH uptake. [3H]PAH uptake showed a non-saturable and a saturable component with similar apparent K m values in brush-border and basolateral membranes. Although one negatively charged PAH molecule exchanges with one doubly negatively charged 2-oxoglutarate molecule the exchange was electroneutral. Probenecid inhibited [3H]PAH/2-oxoglutarate exchange in brush-border and basolateral membrane vesicles with indistinguishable kinetics. We conclude that similar or identical PAH transporters are located in brush-border and basolateral membranes of bovine kidney proximal tubule cells. This arrangement seems species-specific since a Na+ gradient plus 2-oxoglutarate caused concentrative [3H]PAH uptake in brush-border membrane vesicles from bovine, but not from rat kidney.

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  1. Gerhard Burckhardt

    Present address: Zentrum Physiologie und Pathophysiologie, Georg-August-Universität Göttingen, Humboldtallee 23, D-37073, Göttingen, Germany

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  1. Max-Planck-Institut für Biophysik, Kennedyallee 70, W-6000, Frankfurt/Main 70, Germany

    Christiane Schmitt & Gerhard Burckhardt

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  1. Christiane Schmitt
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Schmitt, C., Burckhardt, G. p-Aminohippurate/2-oxoglutarate exchange in bovine renal brush-border and basolateral membrane vesicles. Pflugers Arch. 423, 280–290 (1993). https://doi.org/10.1007/BF00374407

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  • DOI: https://doi.org/10.1007/BF00374407

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