Histochemistry

, Volume 86, Issue 2, pp 207–210 | Cite as

Localization of nucleotide pyrophosphatase in the rat kidney

  • M. Le Hir
  • U. C. Dubach
  • S. Angielski
Article

Summary

Hydrolysis of NAD by a nucleotide pyrophosphatase of renal membrane fractions has been reported previously. The aim of the present study was to localize this enzyme in the rat kidney. Nucleotide pyrophosphatase was assayed in glomeruli, in three parts of the proximal tubule and in four parts of the distal tubule dissected form freezedried sections. Nucleotide pyrophosphatase activity, expressed in μmol·min−1·mg protein−1, ranged between 9.8 and 32.3 in the proximal tubular segments and between 1.1 and 2.7 in the distal tubular segments. It was 3.4 in the glomeruli. The enrichement of the activity during the purification of brush border vesicles was measured. A tenfold higher specific activity was found in the brush border vesicles as compared to the renal cortical homogenates. Thus, most of the renal nucleotide pyrophosphatase appears to be localized in the luminal membrane of the proximal tubule. A permeabilization of the membrane did not increase the activity of brush border vesicles. This indicates that all catalytic sites are accessible at the outer surface of the membrane.

Keywords

Nucleotide Luminal Outer Surface Proximal Tubule Membrane Fraction 

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References

  1. Angielski S, Zielkiewicz J, Dzierko G (1982) Metabolism of NAD by isolated rat renal brush border membranes. Pflügers Arch 395:159–161Google Scholar
  2. Angielski S, Le Hir M, Dubach UC (1983) Transport of adenosine by renal brush border membranes. Pflügers Arch 397:75–77Google Scholar
  3. Biber J, Stieger B, Haase W, Murer H (1981) A high yield preparation for rat kidney brush border membranes. Different behaviour of lysosomal markers. Biochim Biophys Acta 647:169–176Google Scholar
  4. Heinle H, Schmidt U, Wendel A (1977) The activities of the key enzymes of the γ-glutamyl cycle in microdissected segments of the rat nephron. FEBS Lett 73:220–224Google Scholar
  5. Holzbecher J, Ryan DE (1973) The fluorimetric determination of phosphorus with thiamine. Anal Chim Acta 64:147–150Google Scholar
  6. Kesslering K, Siebert G (1964) Eigenschaften einer Dinukleotid Pyrophosphatase aus Rattennieren. Partikeln und Koenzym-Eigenschaften der Spaltprodukte Dihydronicotinamid-Mononukleotid sowie Dihydronicotinamid-Ribosid. Hoppe-Seyler's Z Physiol Chem 337:79–92Google Scholar
  7. Kinne R, Schmitz JE, Kinne-Saffran E (1971) The localization of the Na+−K+-ATPase in the cells of rat kidney cortex. Pflügers Arch 329:191–206Google Scholar
  8. Koseki C, Endou H, Sudo J, Shimada H, Sakai F (1980) Evaluation of nephrotoxic site in rat proximal tubule: intrarenal distribution of three enzymes and effects of mercuric chloride and gentamicin on their excretion into urine. Folia Pharmacol Japon 76:59–69Google Scholar
  9. Kragh-Hansen U, Røigaard-Petersen H, Jacobsen C, Sheikh MI (1984) Renal transport of neutral aminoacids. Tubular localization of Na+-dependent phenylalanine- and glucose-transport systems. Biochem J 220:15–24Google Scholar
  10. Kugler P (1982) On angiotensin—degrading aminopeptidases in the rat kidney. Adv Anat Embryol Cell Biol 76:1–86Google Scholar
  11. Le Hir M, Dubach UV (1982a) Activities of enzymes of the tricarboxylic acid cycle in segments of the rat nephron. Pflügers Arch 395:239–243Google Scholar
  12. Le Hir M, Dubach UC (1982b) The cellular specificity of lectin binding in the kidney. I. A light microscopical study in the rat. Histochemistry 74:521–530Google Scholar
  13. Le Hir M, Angielski S, Dubach UC (1985) Properties of an ecto-5′ nucleotidase of the renal brush border. Renal Physiol 8:321–327Google Scholar
  14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  15. Murer H, Kinne R (1980) The use of isolated membrane vesicles to study epithelial transport processes. J Membr Biol 55:81–95Google Scholar
  16. Naito Y, Lowenstein JM (1981) 5′-Nucleotidase of rat heart. Biochemistry 20:5188–5194Google Scholar
  17. Pfaller W, Rittinger M (1980) Quantitative morphology of the rat kidney. Int J Biochem 12:17–22Google Scholar
  18. Schmidt U, Dubach UC (1971) Quantitative Histochemie am Nephrom. Prog Histochem Cytochem 2:185–298Google Scholar
  19. Spielman WS, Thompson CI (1982) A proposed role for adenosine in the regulation of renal hemodynamics and renine release. Am J Physiol 242:F423-F435Google Scholar
  20. Tenenhouse HS, Chu YL (1982) Hydrolysis of nicotinamide-adenine dinucleotide by purified renal brush-border membrane. Biochem J 204:635–638Google Scholar
  21. Turner RJ, Moran A (1982) Heterogeneity of sodium-dependent d-glucose transport sites along the proximal tubule: evidence from vesicle studies. Am J Physiol 242:F406-F414Google Scholar
  22. Vandewalle A, Wirthensohn G, Heidrich H-G, Guder WG (1981) Distribution of hexokinase and phosphoenolpyrivate carboxykinase along the rabbit nephron. Am J Physiol 240:F492-F500Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • M. Le Hir
    • 1
  • U. C. Dubach
    • 1
  • S. Angielski
    • 2
  1. 1.Department Forschung und Medizinische UniversitätspoliklinikKantonsspitalBaselSwitzerland
  2. 2.Department of Clinical ChemistryMedical AcademyGdanskPoland

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