Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1982 Nov 15;208(2):369–376. doi: 10.1042/bj2080369

The use of a potential-sensitive cyanine dye for studying ion-dependent electrogenic renal transport of organic solutes. Uptake of l-malate and d-malate by luminal-membrane vesicles

Ulrich Kragh-Hansen 1, Karl Evald Jørgensen 1, M Iqbal Sheikh 1
PMCID: PMC1153972  PMID: 7159405

Abstract

The mechanisms of uptake of dicarboxylic acids by rabbit renal luminal-membrane vesicles were studied by the use of filtration and spectrophotometric techniques as described in an accompanying paper [Kragh-Hansen, Jørgensen & Sheikh (1982) Biochem. J. 208, 359–368]. Addition of l- or d-malate to dye-membrane-vesicle suspensions in the presence of Na+ gradients (extravesicular>intravesicular) resulted in spectral curves indicative of depolarization events. The renal uptake of dicarboxylic acids was dependent on the type of Na+-salt anion present and could be correlated with the ability of the anions to penetrate biological membranes (i.e. Cl−>SO42−>gluconate). Identical results were obtained by a filtration technique with Sartorius membrane filters. The results indicate that the dicarboxylic acids are taken up by the membrane vesicles in an electrically positive form (i.e. Na+/substrate coupling ratio 3:1) by an Na+-dependent transport system. This proposal was further supported by spectrophotometric experiments with various ionophores such as valinomycin, gramicidin and nigericin. The absorbance changes associated with simultaneous addition of l- and d-malate and spectrophotometric competition studies revealed that the two isomers are taken up by a common transport system. Spectral changes of the dye induced by addition of increasing concentrations of l- or d-malate indicated that the transport system favours the unphysiological d-form rather than the l-form of malate. Furthermore, it was observed that the affinity of both isomers for the transport system was dependent on the concentration of Na+ in the medium.

Full text

PDF
369

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berner W., Kinne R. Transport of p-aminohippuric acid by plasma membrane vesicles isolated from rat kidney cortex. Pflugers Arch. 1976 Feb 24;361(3):269–277. doi: 10.1007/BF00587292. [DOI] [PubMed] [Google Scholar]
  2. Hopfer U., Nelson K., Perrotto J., Isselbacher K. J. Glucose transport in isolated brush border membrane from rat small intestine. J Biol Chem. 1973 Jan 10;248(1):25–32. [PubMed] [Google Scholar]
  3. Kippen I., Hirayama B., Klinenberg J. R., Wright E. M. Transport of tricarboxylic acid cycle intermediates by membrane vesicles from renal brush border. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3397–3400. doi: 10.1073/pnas.76.7.3397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kook Y. J., Lotspeich W. D. Citrate excretion during intrarenal arterial precursor infusion in the alkalotic dog. Am J Physiol. 1968 Aug;215(2):282–288. doi: 10.1152/ajplegacy.1968.215.2.282. [DOI] [PubMed] [Google Scholar]
  5. Kragh-Hansen U., Jørgensen K. E., Sheikh M. I. The use of potential-sensitive cyanine dye for studying ion-dependent electrogenic renal transport of organic solutes. Spectrophotometric measurements. Biochem J. 1982 Nov 15;208(2):359–368. doi: 10.1042/bj2080359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. LOTSPEICH W. D., WORONKOW S. EXCRETION OF KREBS CYCLE ACIDS IN RELATION TO THE CYCLE'S FUNCTION IN AVIAN KIDNEY. Am J Physiol. 1964 Feb;206:391–402. doi: 10.1152/ajplegacy.1964.206.2.391. [DOI] [PubMed] [Google Scholar]
  7. Ottolenghi P. The reversible delipidation of a solubilized sodium-plus-potassium ion-dependent adenosine triphosphatase from the salt gland of the spiny dogfish. Biochem J. 1975 Oct;151(1):61–66. doi: 10.1042/bj1510061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  9. Runeberg L., Lotspeich W. D. Krebs cycle acid excretion with isotopic split renal function techniques. Am J Physiol. 1966 Aug;211(2):467–475. doi: 10.1152/ajplegacy.1966.211.2.467. [DOI] [PubMed] [Google Scholar]
  10. Sheikh M. I. Renal handling of phenol red. I. A comparative study on the accumulation of phenol red and p-aminohippurate in rabbit kidney tubules in vitro. J Physiol. 1972 Dec;227(2):565–590. doi: 10.1113/jphysiol.1972.sp010048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sheikh M. I. Renal handling of phenol red. II. The mechanism of substituted phenolsulphophthalein (PSP) dye transport in rabbit kidney tubules in vitro. J Physiol. 1976 Mar;256(1):175–195. doi: 10.1113/jphysiol.1976.sp011319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wright S. H., Kippen I., Klinenberg J. R., Wright E. M. Specificity of the transport system for tricarboxylic acid cycle intermediates in renal brush borders. J Membr Biol. 1980 Nov 15;57(1):73–82. doi: 10.1007/BF01868987. [DOI] [PubMed] [Google Scholar]
  13. Wright S. H., Krasne S., Kippen I., Wright E. M. Na+-dependent transport of tricarboxylic acid cycle intermediates by renal brush border membranes. Effects on fluorescence of a potential-sensitive cyanine dye. Biochim Biophys Acta. 1981 Feb 6;640(3):767–778. doi: 10.1016/0005-2736(81)90107-3. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES