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. 1991 Jun;437:495–510. doi: 10.1113/jphysiol.1991.sp018608

The effects of metabolism on Na(+)-K(+)-Cl- co-transport in ferret red cells.

P W Flatman 1
PMCID: PMC1180060  PMID: 1890646

Abstract

1. The effects of altering metabolism on Na(+)-K(+)-Cl- co-transport were studied in ferret red cells. Na(+)-K(+)-Cl- co-transport was measured as the bumetanide-sensitive uptake of 86Rb. 2. Glucose, but not inosine or adenosine, sustained metabolism and maintained cell ATP content ([ATP]i) at the physiological level. [ATP]i could be reduced by prolonged incubation of cells in a substrate-free medium or more quickly by incubating cells with 2-deoxyglucose or with a mixture of iodoacetamide and glucose. 3. Na(+)-K(+)-Cl- co-transport activity was inhibited when [ATP]i was reduced to below 100 mumol (1 cell)-1 by starvation or by treatment with 2-deoxyglucose. However, a unique relationship between [ATP]i and activity could not be found. [ATP]i and the method and time course of ATP depletion all influenced activity. The inhibition of Na(+)-K(+)-Cl- co-transport, caused by reducing [ATP]i could be partially reversed by restoring [ATP]i to normal. 4. Increasing the concentration of intracellular ionized magnesium [( Mg2+]i) did not stimulate co-transport activity in ATP-depleted cells. This contrasts with the substantial stimulation seen in cells with normal [ATP]i. 5. Vanadate stimulated Na(+)-K(+)-Cl- co-transport activity in ATP-depleted cells but not in cells with normal [ATP]i. Fluoride did not affect activity at any [ATP]i. 6. The effects of some sulphydryl reagents on Na(+)-K(+)-Cl- co-transport were also examined. n-Ethylmaleimide (1 mM) inhibited Na(+)-K(+)-Cl- co-transport while it stimulated bumetanide-resistant potassium transport. Dithiothreitol (1 mM) did not affect activity. Iodoacetamide (6 mM) appeared to reduce the inhibition of cotransport activity seen at low [ATP]i but also greatly increased cell fragility. 7. The data suggest that activity of the Na(+)-K(+)-Cl- co-transport system is controlled by a cycle of phosphorylation and dephosphorylation with the phosphorylated form being active. Phosphorylation and transport appear to be almost maximal in ferret red cells with normal [ATP]i. Reduction of [ATP]i may allow changes in phosphatase activity to manifest as changes in transport rate. Differences in the balance between phosphorylation and dephosphorylation may explain tissue-dependent variations in the response of the system to various stimuli.

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Selected References

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

  1. Adragna N. C., Perkins C. M., Lauf P. K. Furosemide-sensitive Na+-K+ cotransport and cellular metabolism in human erythrocytes. Biochim Biophys Acta. 1985 Jan 10;812(1):293–296. doi: 10.1016/0005-2736(85)90551-6. [DOI] [PubMed] [Google Scholar]
  2. Alper S. L., Beam K. G., Greengard P. Hormonal control of Na+-K+ co-transport in turkey erythrocytes. Multiple site phosphorylation of goblin, a high molecular weight protein of the plasma membrane. J Biol Chem. 1980 May 25;255(10):4864–4871. [PubMed] [Google Scholar]
  3. Altamirano A. A., Breitwieser G. E., Russell J. M. Vanadate and fluoride effects on Na+-K+-Cl- cotransport in squid giant axon. Am J Physiol. 1988 Apr;254(4 Pt 1):C582–C586. doi: 10.1152/ajpcell.1988.254.4.C582. [DOI] [PubMed] [Google Scholar]
  4. Bergh C., Kelley S. J., Dunham P. B. K-Cl cotransport in LK sheep erythrocytes: kinetics of stimulation by cell swelling. J Membr Biol. 1990 Aug;117(2):177–188. doi: 10.1007/BF01868684. [DOI] [PubMed] [Google Scholar]
  5. Chipperfield A. R. The (Na+-K+-Cl-) co-transport system. Clin Sci (Lond) 1986 Nov;71(5):465–476. doi: 10.1042/cs0710465. [DOI] [PubMed] [Google Scholar]
  6. Dagher G., Brugnara C., Canessa M. Effect of metabolic depletion on the furosemide-sensitive Na and K fluxes in human red cells. J Membr Biol. 1985;86(2):145–155. doi: 10.1007/BF01870781. [DOI] [PubMed] [Google Scholar]
  7. DiPolo R., Beaugé L. In squid axons, ATP modulates Na+-Ca2+ exchange by a Ca2+i-dependent phosphorylation. Biochim Biophys Acta. 1987 Mar 12;897(3):347–354. doi: 10.1016/0005-2736(87)90432-9. [DOI] [PubMed] [Google Scholar]
  8. Duhm J., Göbel B. O. Role of the furosemide-sensitive Na+/K+ transport system in determining the steady-state Na+ and K+ content and volume of human erythrocytes in vitro and in vivo. J Membr Biol. 1984;77(3):243–254. doi: 10.1007/BF01870572. [DOI] [PubMed] [Google Scholar]
  9. Flatman P. W., Lew V. L. Magnesium buffering in intact human red blood cells measured using the ionophore A23187. J Physiol. 1980 Aug;305:13–30. doi: 10.1113/jphysiol.1980.sp013346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Flatman P. W. Sodium and potassium transport in ferret red cells. J Physiol. 1983 Aug;341:545–557. doi: 10.1113/jphysiol.1983.sp014823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Flatman P. W. The effects of calcium on potassium transport in ferret red cells. J Physiol. 1987 May;386:407–423. doi: 10.1113/jphysiol.1987.sp016541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Flatman P. W. The effects of magnesium on potassium transport in ferret red cells. J Physiol. 1988 Mar;397:471–487. doi: 10.1113/jphysiol.1988.sp017013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Geck P., Pietrzyk C., Burckhardt B. C., Pfeiffer B., Heinz E. Electrically silent cotransport on Na+, K+ and Cl- in Ehrlich cells. Biochim Biophys Acta. 1980 Aug 4;600(2):432–447. doi: 10.1016/0005-2736(80)90446-0. [DOI] [PubMed] [Google Scholar]
  14. Haas M. Properties and diversity of (Na-K-Cl) cotransporters. Annu Rev Physiol. 1989;51:443–457. doi: 10.1146/annurev.ph.51.030189.002303. [DOI] [PubMed] [Google Scholar]
  15. Haas M., Schmidt W. F., 3rd, McManus T. J. Catecholamine-stimulated ion transport in duck red cells. Gradient effects in electrically neutral [Na + K + 2Cl] Co-transport. J Gen Physiol. 1982 Jul;80(1):125–147. doi: 10.1085/jgp.80.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hall A. C., Ellory J. C. Measurement and stoichiometry of bumetanide-sensitive (2Na:1K:3Cl) cotransport in ferret red cells. J Membr Biol. 1985;85(3):205–213. doi: 10.1007/BF01871515. [DOI] [PubMed] [Google Scholar]
  17. Jennings M. L., al-Rohil N. Kinetics of activation and inactivation of swelling-stimulated K+/Cl- transport. The volume-sensitive parameter is the rate constant for inactivation. J Gen Physiol. 1990 Jun;95(6):1021–1040. doi: 10.1085/jgp.95.6.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kim H. D., Tsai Y. S., Franklin C. C., Turner J. T. Characterization of Na+/K+/Cl- cotransport in cultured HT29 human colonic adenocarcinoma cells. Biochim Biophys Acta. 1988 Dec 22;946(2):397–404. doi: 10.1016/0005-2736(88)90415-4. [DOI] [PubMed] [Google Scholar]
  19. Lauf P. K. K+:Cl- cotransport: sulfhydryls, divalent cations, and the mechanism of volume activation in a red cell. J Membr Biol. 1985;88(1):1–13. doi: 10.1007/BF01871208. [DOI] [PubMed] [Google Scholar]
  20. Lauf P. K. Thiol-dependent passive K/Cl transport in sheep red cells: VII. Volume-independent freezing by iodoacetamide, and sulfhydryl group heterogeneity. J Membr Biol. 1987;98(3):237–246. doi: 10.1007/BF01871186. [DOI] [PubMed] [Google Scholar]
  21. Lew V. L. On the ATP dependence of the Ca 2+ -induced increase in K + permeability observed in human red cells. Biochim Biophys Acta. 1971 Jun 1;233(3):827–830. doi: 10.1016/0005-2736(71)90185-4. [DOI] [PubMed] [Google Scholar]
  22. Li H. C. Phosphoprotein phosphatases. Curr Top Cell Regul. 1982;21:129–174. [PubMed] [Google Scholar]
  23. Macdonald T. L., Martin R. B. Aluminum ion in biological systems. Trends Biochem Sci. 1988 Jan;13(1):15–19. doi: 10.1016/0968-0004(88)90012-6. [DOI] [PubMed] [Google Scholar]
  24. Palfrey H. C., Rao M. C. Na/K/Cl co-transport and its regulation. J Exp Biol. 1983 Sep;106:43–54. doi: 10.1242/jeb.106.1.43. [DOI] [PubMed] [Google Scholar]
  25. Rindler M. J., McRoberts J. A., Saier M. H., Jr (Na+,K+)-cotransport in the Madin-Darby canine kidney cell line. Kinetic characterization of the interaction between Na+ and K+. J Biol Chem. 1982 Mar 10;257(5):2254–2259. [PubMed] [Google Scholar]
  26. Russell J. M. Anion transport mechanisms in neurons. Ann N Y Acad Sci. 1980;341:510–523. doi: 10.1111/j.1749-6632.1980.tb47195.x. [DOI] [PubMed] [Google Scholar]
  27. SOLS A., CRANE R. K. Substrate specificity of brain hexokinase. J Biol Chem. 1954 Oct;210(2):581–595. [PubMed] [Google Scholar]
  28. Saier M. H., Jr, Boyden D. A. Mechanism, regulation and physiological significance of the loop diuretic-sensitive NaCl/KCl symport system in animal cells. Mol Cell Biochem. 1984;59(1-2):11–32. doi: 10.1007/BF00231303. [DOI] [PubMed] [Google Scholar]
  29. Swarup G., Cohen S., Garbers D. L. Inhibition of membrane phosphotyrosyl-protein phosphatase activity by vanadate. Biochem Biophys Res Commun. 1982 Aug;107(3):1104–1109. doi: 10.1016/0006-291x(82)90635-0. [DOI] [PubMed] [Google Scholar]
  30. Ueberschär S., Bakker-Grunwald T. Effects of ATP and cyclic AMP on the (Na+ + K+ + 2Cl-)-cotransport system in turkey erythrocytes. Biochim Biophys Acta. 1985 Aug 27;818(2):260–266. doi: 10.1016/0005-2736(85)90566-8. [DOI] [PubMed] [Google Scholar]
  31. Whittam R., Wiley J. S. Some aspects of adenosine triphosphate synthesis from adenine and adenosine in human red blood cells. J Physiol. 1968 Dec;199(2):485–494. doi: 10.1113/jphysiol.1968.sp008664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wiater L. A., Dunham P. B. Passive transport of K+ and Na+ in human red blood cells: sulfhydryl binding agents and furosemide. Am J Physiol. 1983 Nov;245(5 Pt 1):C348–C356. doi: 10.1152/ajpcell.1983.245.5.C348. [DOI] [PubMed] [Google Scholar]

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