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Protein kinase C consensus sites and the regulation of renal Na/Pi-cotransport (NaPi-2) expressed in XENOPUS laevis oocytes

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  • Molecular and Cellular Physiology
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Abstract

Renal brush border membrane sodium/phosphate (Na/Pi)-cotransport activity is inhibited by hormonal mechanisms involving activation of protein kinases A and C. The recently cloned rat renal Na/Pi cotransporter (NaPi-2) contains several protein kinase C but no protein kinase A consensus sites [17, 20]. In the present study we have expressed wild type and polymutant (protein kinase C consensus sites removed) NaPi-2-transporters in Xenopus laevis oocytes. The expression of transport function as well as the basic transport properties were unaffected by the removal of the consensus sites. Pharmacological activation of protein kinase C with phorbol 12,13-didecanoate (PDD) led to a time-dependent inhibition of expressed wild type Na/Picotransport function; simultaneous exposure to staurosporine (0.3) prevented the PDD induced (50 nM) inhibition. The kinase-C-mediated inhibition was not prevented by the removal of the protein kinase C consensus sites. Pharmacological activation of protein kinase A (dibutyryl adenosine 3′:5′:cyclic monophosphate (cAMP)/forskolin) had no effect on wild type NaPi-2-induced oocyte Na/Pi cotransport. It is concluded that the protein-kinase-C-mediated regulation of expressed Na/Pi-cotransport does not involve the predicted consensus sites. The involvement of “cryptic” phosphorylation sites and/or of a phosphorylated “regulatory” protein is discussed.

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References

  1. Beguin P, Beggah AT, Chibalin AV, Burgener-Kairuz P, Jaisser F, Mathews PM, Rossier BC, Cotecchia S, Geering K (1994) Phosphorylation of the Na, K-ATPase α-subunit by protein kinase A and C in vitro and in intact cells. J Biol Chem 269:24437–24445

    Google Scholar 

  2. Berndt TJ, Knox FG (1992) Renal regulation of phosphate excretion. In:Seldin DW, Giebisch G (eds) The kidney, physiology and pathophysiology, (2nd edn) Raven, New York, pp 2511–2532

    Google Scholar 

  3. Blumenthal EM, Kaczmarek LK (1992) Modulation by cAMP of a slowly activating potassium channel expressed in Xenopus oocytes. J Neurosci 12:290–296

    Google Scholar 

  4. Busch AE, Varnum MD, North RA, Adelman JP (1992) An amino acid mutation in a potassium channel that prevents inhibition by protein kinase C. Science 255:1705–1707

    Google Scholar 

  5. Busch AE, Kavanaugh MP, Varnum MD, Adelman JP, North RA (1992) Regulation of the slowly activating, voltage-dependent potassium channel expressed in Xenopus oocytes. J Physiol (Lond) 450:491–502

    Google Scholar 

  6. Busch A, Waldegger S, Herzer T, Biber J, Markovich D, Hayes G, Murer H, Lang F (1994) Electrophysiological analysis of Na/Pi cotransport mediated by a transporter cloned from rat kidney and expressed in Xenopus oocytes. Proc Natl Acad Sci USA 91:8205–8208

    Google Scholar 

  7. Chang XB, Tabcharani JA, Hou YX, Jensen TJ, Kartner N, Alon N, Hanrahan JW, Riordan JR (1993) Protein kinase A (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites. J Biol Chem 268:11304–11311

    Google Scholar 

  8. Custer M, Lötscher M, Biber J, Murer H, Kaissling B (1994) Expression of Na/Pi-cotransport (NaPi-2) in rat kidney: localization by RT-PCR and immunohistochemistry. Am J Physiol 266:F767-F774

    Google Scholar 

  9. Dennis V (1992) Phosphate homeostasis. In: Windhager EE (ed) Handbook of physiology. Oxford University Press, New York, pp 1785–1815

    Google Scholar 

  10. Dousa TP, Duarte CG, Knox FG (1976) Effect of colchicine on urinary phosphate and regulation by parathyroid hormone. Am J Physiol 231:61–65

    Google Scholar 

  11. Dunlay R, Hruska K (1990) PTH receptor coupling to phospolipase C is an alternate pathway of signal transduction in bone and kidney. Am J Physiol 258:F223-F231

    Google Scholar 

  12. Hammerman MR (1986) Phosphate transport across renal proximal tubular cell membranes. Am J Physiol 251:F385-F398

    Google Scholar 

  13. Hayes G, Busch A, Lötscher M, Waldegger S, Lang F, Verrey F, Biber J, Murer H (1994) Role of N-linked glycosylation in rat renal Na/Picotransport. J Biol Chem 269:24143–24149

    Google Scholar 

  14. Kempson SA, Helmle-Kolb C, Abraham MI, Murer H (1990) Parathyroid hormone action on phosphate transport is inhibited by high osmolality. Am J Physiol 258:F1336-F1344

    Google Scholar 

  15. Kempson SA, Lötscher M, Kaissling B, Biber J, Murer H, Levi M (1995) Parathyroid hormone action and phosphate transporter mRNA and protein in rat renal proximal tubules. Am J Physiol 268:F784-F791

    Google Scholar 

  16. Levi M, Lötscher M, Sorribas V, Custer M, Arar M, Kaissling B, Murer H, Biber J (1994) Cellular mechanisms of acute and chronic adaptation of rat renal Pi transporter to alterations in dietary Pi. Am J Physiol 267:F900-F908

    Google Scholar 

  17. Magagnin S, Werner A, Markovich D, Sorribas V, Stange G, Biber J, Murer H (1993) Expression cloning of human rat renal cortex Na/Pi cotransport. Proc Natl Acad Sci USA 90:5979–5983

    Google Scholar 

  18. Murai T, Kakizuka A, Takumi T, Ohkubo H, Nakanishi S (1989) Molecular cloning and sequence analysis of human genomic DNA encoding a novel membrane protein which exhibits a slowly activating potassium channel activity. Biochem Biophys Res Commun 161:176–181

    Google Scholar 

  19. Murer H, Biber J (1992) Renal tubular phosphate transport: cellular mechanisms. In: Seldin DW, Giebisch G (eds) The kidney: physiology and pathophysiology, (2nd edn). Raven, New York, pp 2481–2509

    Google Scholar 

  20. Murer H, Biber J (1994) Renal sodium-phosphate cotransport. Curr Opinion Nephrol Hypertens 3:504–510

    Google Scholar 

  21. Murer H, Werner A, Reshkin S, Wuarin F, Biber J (1991) Cellular mechanisms in proximal tubular reabsorption of inorganic phosphate. Am J Physiol 260:C885-C899

    Google Scholar 

  22. Takumi T, Otikubo H, Nakanishi S (1988) Cloning of a membrane protein that induces a slow voltage-gated potassium current. Science 242:1042–1045

    Google Scholar 

  23. Vasilets LA, Schmalzing G, Madefessel K, Haase W, Schwarz W (1990) Activation of protein kinase C by phorbol ester induces downregulation of the Na+/K+-ATPase in oocytes of Xenopus laevis. J Membr Biol 118:131–142

    Google Scholar 

  24. Weinman EJ, Steplock D, Shenolikar S (1993) cAMP-mediated inhibition of the renal brush border membrane Na+-H+ exchanger requires a dissociable phosphoprotein cofactor. J Clin Invest 92:1781–1786

    Google Scholar 

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Authors and Affiliations

  1. Institute of Physiology, University of Zürich, Winterthurerstrasse 190, CH-8057, Zürich, Switzerland

    Gillian Hayes, Jürg Biber & Heini Murer

  2. Institute of Physiology I, Eberhard-Karls Universität, Gmelinstrasse 5, D-72076, Tübingen, Germany

    Andreas E. Busch & Florian Lang

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  1. Gillian Hayes
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  2. Andreas E. Busch
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  3. Florian Lang
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  4. Jürg Biber
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  5. Heini Murer
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Hayes, G., Busch, A.E., Lang, F. et al. Protein kinase C consensus sites and the regulation of renal Na/Pi-cotransport (NaPi-2) expressed in XENOPUS laevis oocytes. Pflugers Arch. 430, 819–824 (1995). https://doi.org/10.1007/BF00386181

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  • DOI: https://doi.org/10.1007/BF00386181

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