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Effect of SH-, NH2- and COOH-site group reagents on the transport processes in the proximal convolution of the rat kidney

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Summary

The effects of site group reagents were tested on the following transport processes of the proximal convolution. Isotonic Na+ absorption, evaluated by the shrinking droplet procedure, histidine and glucose transport, evaluated by measuring the respective transtubular concentration difference at zero substance and water net flux. The test substances were applied either by continuous microperfusion of the peritubular capillaries or by luminal perfusion prior to the transport tests or by addition to the luminal test solution.

The SH reagents (0.2 mM) N-ethylmaleimide,p-chloromercuribenzoate (pCMB) 3,6-bis-(acetatomercurimethyl)dioxane and Mersalyl (Salyrgane) caused 50% inhibition of the isotonic Na+ absorption in approximately 1.5 min when applied to the capillary perfusate. The same effect was reached in 2–3 min by 0.2 mMp-chloromercuriphenylsulfonate, benzamido-4-iodo-acetylstilbene-2,5-disulfonate and 2,2′-dihydroperoxy-2,2′-dibutylperoxide. However, the large molecular SH reagentspCMB-dextran T10 and benzoxanthene-3,4-dicarboxylic-N-iodoacetyloligoprolyl-2-aminoethylimid, did not inhibit the isotonic Na+ absorption. If an inhibitory effect was observed on the Na+ transport its onset was faster, when the substance was applied from the blood site than when it was given from the tubular lumen. Because SH reagents inhibit the isotonic Na transport faster when applied from the blood side, and because SH reagents with MW up to 690 are inhibitory whereas larger ones with MW over 1700 are not, it seems that they exert their inhibitory action on SH groups located a) predominantly on the blood side and b) deep within the membrane and not at the surface.

Histidine- and glucose transport was inhibited only when the sodium transport was inhibited considerably.

The oxygen consumption of teased kidney slices is not inhibited by 0.2 mMpCMB or Mersalyl within 10 min, but it is inhibited considerably by 1 mM of these substances in the same period of incubation time.

The COOH reagents N,N′-carbonyl-diimidazole and N-ethyl-N′-(3-dimethyl-aminopropyl)carbodiimid (10 mM) and the NH2 reagents 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid, 2 Na+ (SITS) (1 mM) as well as danslychloride (applied from the lumen at 5 mM in paraffin oil) did not inhibit the isotonic Na+ absorption.

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References

  1. Bode, F., Baumann, K., Frasch, W., Kinne, R.: Die Bindung von Phlorrhizin an die Bürstensaumfraktion der Rattenniere. Pflügers Arch.315, 53–65 (1970)

    Google Scholar 

  2. Borghgraef, R. R. M., Pitts, R. F.: The distribution of chlormerodrin (neohydrin) in tissues of the rat and dog. J. clin. Invest.35, 31–37 (1956)

    Google Scholar 

  3. Bowman, F. J., Landon, E. J.: Organic mercurials and net movements of potassium in rat kidney slices. Amer. J. Physiol.213, 1209–1217 (1967)

    Google Scholar 

  4. Bretscher, M. S.: Human erythrocyte membranes: specific labelling of surface proteins. J. molec. Biol.58, 775–781 (1971)

    Google Scholar 

  5. Clarkson, T. W., Greenwood, M.: The mechanism of action of mercurial diuretics in rats; the renal metabolism of p-chloromercuribenzoate and its effect on urinary excretion. Brit. J. Pharmacol.26, 50–55 (1966)

    Google Scholar 

  6. Eldjarn, L., Jellum, E.: Organomercurial-polysaccharide, a chromatographic material for the separation and isolation of SH-proteins. Acta chem. scand.17, 2610–2621 (1963)

    Google Scholar 

  7. Frasch, W., Frohnert, P. P., Bode, F., Baumann, K., Kinne, R.: Competitive inhibition of phlorizin binding byd-glucose and the influence of sodium: a study on isolated brush border membranes of rat kidney. Pflügers Arch.320, 265–284 (1970)

    Google Scholar 

  8. Frömter, E., Diamond, J. M.: Route of passive ion permeation in epithelia. Nature New Biol.235, 9–13 (1972)

    Google Scholar 

  9. Frömter, E., Müller, C. W., Wick, T.: Permeability properties of the proximal tubular epithelium of the rat kidney studied with electrophysiological methods. In: Electrophysiology of epithelial cells, pp. 119–146, G. Giebisch, Ed. Stuttgart-New York: F. K. Schattauer 1971

    Google Scholar 

  10. Fülgraff, G.: Effects of diuretics on the relation between oxygen consumption and sodium transport. Proc. 4th Int. Congr. Nephrol., Stockholm 1969, vol. 2, pp. 119–126. Basel-München-New York: Karger 1970

    Google Scholar 

  11. Gertz, K. H.: Transtubuläre Natriumchloridflüsse und Permeabilität für Nichtelektrolyte im proximalen und distalen Konvolut der Rattenniere. Pflügers Arch. ges. Physiol.276, 336–356 (1963)

    Google Scholar 

  12. Györy, A. Z.: Reexamination of the split oil droplet method as applied to kidney tubules. Pflügers Arch.324, 328–343 (1971)

    Google Scholar 

  13. Heidenreich, O.: Quecksilberhaltige Diuretica. In: Handbook of experimental Pharmacology, vol. XXIV: Diuretica, pp. 62–194. Berlin-Heidelberg-New York: Springer 1969

    Google Scholar 

  14. Heidrich, H. G., Kinne, R., Kinne-Saffran, E., Hannig, K.: The polarity of the proximal tubule cell in rat kidney. Different surface charges for the brush border microvilli and plasma membranes from the basal infoldings. J. Cell Biol.54, 232–245 (1972)

    Google Scholar 

  15. Hillman, R. E., Rosenberg, L. E.: Aminoacid transport by isolated mammalian renal tubules. III. Binding of — proline by proximal tubule membranes. Biochim. biophys. Acta (Amst.)211, 318–326 (1970)

    Google Scholar 

  16. Kleinzeller, A., Cort, J. H.: The mechanism of action of mercurial preparations on transport processes and the role of thiol groups in the cell membrane of renal tubular cells. Biochem. J.67, 15–24 (1957)

    Google Scholar 

  17. Kleinzeller, A., Janacek, K.: The binding of mercury (203Hg) by animal tissue in vitro. Physiol. bohemoslov.11, 285–293 (1962)

    Google Scholar 

  18. Knauf, P. A., Rothstein, A.: Chemical modification of membranes. I. Effects of sulfhydryl and amino reactive reagents on anion and cation permeability of the human red blood cell. J. gen. Physiol.58, 190–210 (1971).

    Google Scholar 

  19. —: Chemical modification of membranes. II. Permeation path for sulfhydryl agents. J. gen. Physiol.58, 211–223 (1971)

    Google Scholar 

  20. Kotaki, A., Naoi, M., Yagi, K.: A diaminostilbene dye as a hydrophobic probe for proteins. Biochim. biophys. Acta (Amst.)229, 547–556 (1971)

    Google Scholar 

  21. Lengsfeld, A., Hasselbach, W.: Struktur und chemische Asymmetrie von Nierenmembranen. Elektronenmikroskopische Untersuchungen an verschiedenen Membranfraktionen des äußeren Nierencortex (Schwein). Histochemie27, 253–276 (1971)

    Google Scholar 

  22. Means, G. E., Feeney, R. E.: Chemical modification of proteins. San Francisco-Cambridge-London-Amsterdam: Holden Day Inc. 1971

    Google Scholar 

  23. Milas, N. A., Golubović, A.: Organic peroxides XXV. Preparation, separation and identification of peroxides derived from methyl, ethyl, ketone and hydrogen peroxide. J. Amer. chem. Soc.81, 5824–5826 (1959)

    Google Scholar 

  24. Nechay, B., Palmer, R. F., Chinoy, D. A., Possey, V. A.: The problem of Na+K+ ATPase as receptor for diuretic action of mercurials and ethacrynic acid. J. Pharmacol. exp. Ther.157, 599–617 (1967)

    Google Scholar 

  25. Newey, H., Sanford, P. A., Smyth, D. H.: Some effects of ouabain and potassium on transport and metabolism in rat small intestine. J. Physiol. (Lond.)194, 237–248 (1968)

    Google Scholar 

  26. Ohta, H., Matsumoto, J., Nagano, K., Fujita, M., Nakao, M.: The inhibition of Na+K+ activated adenosine-triphosphatase by a large molecule derivative of p-chloromercuribenzoic acid at the outer surface of human red cell. Biochem. biophys. Res. Commun.42, 1127–1133 (1971)

    Google Scholar 

  27. Rambourg, A., Leblond, C. P.: Electron microscope observations on the carbohydrate-rich cell coat present at the surface of cells in the rat. J. Cell. Biol.32, 27–53 (1967)

    Google Scholar 

  28. Salzer, M.: Synthese und Modellreaktionen von Protein markierenden Fluoreszenzfarbstoffen. Chem. Diplomarbeit an der Johann Wolfgang von Goethe-Universität Frankfurt (1972)

  29. Sato, K.: Inhibition of respiration in the isolated eccrine sweat gland by ethacrynic acid. Pflügers Arch.341, 233–242 (1973)

    Google Scholar 

  30. Silverman, M., Aganon, M. A., Chinard, F. P.:d-glucose interactions with renal tubule cell surfaces. Amer. J. Physiol.218, 735–742 (1970)

    Google Scholar 

  31. Simon, B., Zimmerschied, G., Kinne-Saffran, E., Kinne, R.: Properties of a synthetic plasma membrane marker: Fluorescent-Mercury-Dextran. Biochim. biophys. Acta (Amst.) (in press)

  32. Skou, J. C., Hilberg, C.: The effect of sulfhydryl blocking reagents and of urea on the (Na++K+)-activated enzyme system. Biochim. biophys. Acta (Amst.)110, 359–369 (1965)

    Google Scholar 

  33. Van Stevenick, J., Weed, R. I., Rothstein, A.: Localization of erythrocyte membrane sulfhydryl groups essential for glucose transport. J. gen. Physiol.48, 617–632 (1965)

    Google Scholar 

  34. Taylor, C. B.: The effect of mercurial diuretics on adenosine triphosphatase of rabbit kideny in vitro. Biochem. Pharmacol.12, 539–550 (1963)

    Google Scholar 

  35. Thomas, L., Kinne, R.: Studies on the arrangement of glucose sensitive phlorizin binding site in the microvilli of isolated rat kidney brush border. FEBS Letters25, 242–244 (1972)

    Google Scholar 

  36. Ullrich, K. J.: Permeability characteristics of the mammalian nephron. Handbook of Physiology, Section 8: Renal physiology, pp. 377–398, J. Orloff and R. W. Berliner, Eds. Amer. Physiol. Society 1973.

  37. Ullrich, K. J., Frömter, E., Baumann, K.: Micropuncture and microanalysis in kidney physiology. In: Laboratory techniques in membrane biophysics, pp. 106–129, H. Passow and R. Stämpfli, Eds. Berlin-Heidelberg-New York: Springer 1969

    Google Scholar 

  38. Ullrich, K. J., Rumrich, G., Klöss, S.: Sodium dependence of the transtubular transport of glucose and aminoacid in the proximal tubule of the rat kidney. Pflügers Arch.339, Suppl. R47 (1973)

    Google Scholar 

  39. Ullrich, K. J., Sato, K., Rumrich, G., Klöss, S.: coupling of the transport processes across the brush border of the proximal renal tubule, pp. 560–571. In: Alfred Benzon Symposium V: Transport mechanisms in epithelia, H. H. Ussing and N. A. Thorn, Eds. Copenhagen: Munksgaard and New York: Academic Press 1973

    Google Scholar 

  40. Wallach, D. F. H.: The dispositions of proteins in the plasma membranes of animal cells: Analytical approaches using controlled peptidolysis and protein labels. Biochim. biophys. Acta (Amst.)265, 61–83 (1972)

    Google Scholar 

  41. Weed, R. I., Berg, G. G.: Membrane sulfhydryl groups and ATPase. Fed. Proc.22, A 213 (1963), p. 23

    Google Scholar 

  42. Weeden, R. P., Weiner, B.: The distribution of p-aminohippuric acid in rat kidney slices. I. Tubular localization. Kidney Intern.3, 205–213 (1973)

    Google Scholar 

  43. Whittam, R., Willis, J. S.: Ion movements and oxygen consumption in kidney cortex slices. J. Physiol. (Lond.)168, 158–177 (1963)

    Google Scholar 

  44. Wizemann, V., Schulz, I., Simon, B.: SH-groups on the surface of pancreas cells involved in secretion stimulation and glucose-mediated secretion. Biochim. biophys. Acta (Amst.)307, 366–371 (1973)

    Google Scholar 

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Authors and Affiliations

  1. Max-Planck-Institut für Biophysik, Frankfurt/Main, Germany

    K. J. Ullrich, H. Fasold, S. Klöss, G. Rumrich, M. Salzer, K. Sato, B. Simon & J. X. de Vries

  2. Institut für Biochemie der Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany

    K. J. Ullrich, H. Fasold, S. Klöss, G. Rumrich, M. Salzer, K. Sato, B. Simon & J. X. de Vries

  3. Max-Planck-Institut für Medizinische Forschung, Heidelberg, Germany

    K. J. Ullrich, H. Fasold, S. Klöss, G. Rumrich, M. Salzer, K. Sato, B. Simon & J. X. de Vries

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Ullrich, K.J., Fasold, H., Klöss, S. et al. Effect of SH-, NH2- and COOH-site group reagents on the transport processes in the proximal convolution of the rat kidney. Pflugers Arch. 344, 51–68 (1973). https://doi.org/10.1007/BF00587441

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