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Monovalent cation leaks in human red cells caused by single amino-acid substitutions in the transport domain of the band 3 chloride-bicarbonate exchanger, AE1

Nature Genetics volume 37, pages 1258–1263 (2005)Cite this article

Abstract

We identified 11 human pedigrees with dominantly inherited hemolytic anemias in both the hereditary stomatocytosis and spherocytosis classes. Affected individuals in these families had an increase in membrane permeability to Na and K that is particularly marked at 0 °C. We found that disease in these pedigrees was associated with a series of single amino-acid substitutions in the intramembrane domain of the erythrocyte band 3 anion exchanger, AE1. Anion movements were reduced in the abnormal red cells. The 'leak' cation fluxes were inhibited by SITS, dipyridamole and NS1652, chemically diverse inhibitors of band 3. Expression of the mutated genes in Xenopus laevis oocytes induced abnormal Na and K fluxes in the oocytes, and the induced Cl transport was low. These data are consistent with the suggestion that the substitutions convert the protein from an anion exchanger into an unregulated cation channel.

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Figure 1: Temperature dependence of leak cation flux in different individuals.
Figure 2: Effect of band 3 inhibitors on cation leaks in different pedigrees.
Figure 3: Anion transport in normal and abnormal cells.
Figure 4: Heterologous expression in X. laevis oocytes.
Figure 5: Assessment of expression of mutant and wild-type band 3 in two individuals with CHC with band 3 Memphis.

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References

  1. Stewart, G.W. Hemolytic disease due to membrane ion channel disorders. Curr. Opin. Hematol. 11, 244–250 (2004).

    Article  CAS  Google Scholar 

  2. Jarolim, P. et al. Characterization of 13 novel band 3 gene defects in hereditary spherocytosis with band 3 deficiency. Blood 88, 4366–4374 (1996).

    CAS  PubMed  Google Scholar 

  3. Dhermy, D. et al. Heterogenous band 3 deficiency in hereditary spherocytosis related to different band 3 gene defects. Br. J. Haematol. 98, 32–40 (1997).

    Article  CAS  Google Scholar 

  4. Yannoukakos, D. et al. Human erythrocyte Band 3 polymorphism (Band 3 Memphis): characterization of the structural modification (Lys56-Glu) by protein chemistry methods. Blood 78, 1117–1120 (1991).

    CAS  PubMed  Google Scholar 

  5. Knauf, P.A. & Pal, P. Band 3 mediated transport. in Red Cell Membrane Transport in Health and Disease (eds. Bernhardt, I. & Ellory, J.C.) 253–301 (Springer, Berlin, 2003).

    Chapter  Google Scholar 

  6. Bennekou, P., Pedersen, O., Moller, A. & Christophersen, P. Volume control in sickle cells is facilitated by the novel anion conductance inhibitor NS1652. Blood 95, 1842–1848 (2000).

    CAS  PubMed  Google Scholar 

  7. Bruce, L.J., Unwin, R.J., Wrong, O. & Tanner, M.J. The association between familial distal renal tubular acidosis and mutations in the red cell anion exchanger (band 3, AE1) gene. Biochem. Cell Biol. 76, 723–728 (1998).

    Article  CAS  Google Scholar 

  8. Haines, P. et al. Two further British families with the 'cryohydrocytosis' form of hereditary stomatocytosis. Br. J. Haematol. 113, 932–937 (2001).

    Article  CAS  Google Scholar 

  9. Eber, S. & Lux, S.E. Hereditary spherocytosis–defects in proteins that connect the membrane skeleton to the lipid bilayer. Semin. Hematol. 41, 118–141 (2004).

    Article  CAS  Google Scholar 

  10. Coles, S. et al. Two British families with variants on the 'cryohydrocytosis' form of hereditary stomatocytosis. Br. J. Haematol. 105, 1055–1065 (1999).

    Article  CAS  Google Scholar 

  11. Bruce, L.J. et al. South-east asian ovalocytic (SAO) erythrocytes have a cold sensitive cation leak: implications for in vitro studies on stored SAO red cells. Biochim. Biophys. Acta 1416, 258–270 (1999).

    Article  CAS  Google Scholar 

  12. Guizouarn, H., Gabillat, N., Motais, R. & Borgese, F. Multiple transport functions of a red blood cell anion exchanger, tAE1: its role in cell volume regulation. J. Physiol. (Lond.) 535, 497–506 (2001).

    Article  CAS  Google Scholar 

  13. DeFelice, L.J. Transporter structure and mechanism. Trends Neurosci. 27, 352–359 (2004).

    Article  CAS  Google Scholar 

  14. Tosteson, M.T., Halperin, J.A., Kishi, Y. & Tosteson, D.C. Palytoxin induces an increase in the cation conductance of red cells. J. Gen. Physiol. 98, 969–985 (1991).

    Article  CAS  Google Scholar 

  15. Meredith, D. Site-directed mutation of arginine 282 to glutamate uncouples the movement of peptides and protons by the rabbit proton-peptide cotransporter PepT1. J. Biol. Chem. 279, 15795–15798 (2004).

    Article  CAS  Google Scholar 

  16. Accardi, A. & Miller, C. Secondary active transport mediated by a prokaryotic homologue of ClC Cl- channels. Nature 427, 803–807 (2004).

    Article  CAS  Google Scholar 

  17. Jones, G.S. & Knauf, P.A. Mechanism of the increase in cation permeability of human erythrocytes in low-chloride media. Involvement of the anion transport protein capnophorin. J. Gen. Physiol. 86, 721–738 (1985).

    Article  CAS  Google Scholar 

  18. Johnson, R.M. & Tang, K. DIDS inhibition of deformation-induced cation flux in human erythrocytes. Biochim. Biophys. Acta 1148, 7–14 (1993).

    Article  CAS  Google Scholar 

  19. Joiner, C.H. Deoxygenation-induced cation fluxes in sickle cells: II. Inhibition by stilbene disulfonates. Blood 76, 212–220 (1990).

    CAS  PubMed  Google Scholar 

  20. Joiner, C.H. et al. Dipyridamole inhibits sickling-induced cation fluxes in sickle red blood cells. Blood 97, 3976–3983 (2001).

    Article  CAS  Google Scholar 

  21. Stuart, M.J. & Nagel, R.L. Sickle-cell disease. Lancet 364, 1343–1360 (2004).

    Article  Google Scholar 

  22. Lew, V.L. & Bookchin, R.M. Ion transport pathology in the mechanism of sickle cell dehydration. Physiol. Rev. 85, 179–200 (2005).

    Article  CAS  Google Scholar 

  23. Fricke, B. et al. Four new cases of stomatin-deficient hereditary stomatocytosis syndrome: association of the stomatin-deficient cryohydrocytosis variant with neurological dysfunction. Br. J. Haematol. 125, 796–803 (2004).

    Article  CAS  Google Scholar 

  24. Coles, S.E., Ho, M.M., Chetty, M.C., Nicolaou, A. & Stewart, G.W. Hereditary stomatocytosis with marked pseudohyperkalaemia. Br. J. Haematol. 104, 275–283 (1999).

    Article  CAS  Google Scholar 

  25. Gore, D.M. et al. Four pedigrees of the cation-leaky hereditary stomatocytosis class presenting with pseudohyperkalaemia. Novel profile of temperature dependence of Na+-K+ leak in a xerocytic form. Br. J. Haematol. 125, 521–527 (2004).

    Article  CAS  Google Scholar 

  26. Gore, D.M., Chetty, M.C., Coles, S.C., Gover, P.M. & Stewart, G.W. Hereditary spherocytosis with a low temperature Na/K leak and thrombosis. Br. J. Haematol. 117 Suppl. 1, 9–10 (2002).

    Google Scholar 

  27. Stewart, G.W. et al. Thrombo-embolic disease after splenectomy for hereditary stomatocytosis. Br. J. Haematol. 93, 303–310 (1996).

    Article  CAS  Google Scholar 

  28. Fricke, B. et al. The 'stomatin' gene and protein in overhydrated hereditary stomatocytosis. Blood 102, 2268–2277 (2003).

    Article  CAS  Google Scholar 

  29. Bruce, L.J., Anstee, D.J., Spring, F.A. & Tanner, M.J. Band 3 Memphis variant II. Altered stilbene disulfonate binding and the Diego (Dia) blood group antigen are associated with the human erythrocyte band 3 mutation Pro854 → Leu. J. Biol. Chem. 269, 16155–16158 (1994).

    CAS  PubMed  Google Scholar 

  30. Popov, M., Li, J. & Reithmeier, R.A. Transmembrane folding of the human erythrocyte anion exchanger (AE1, Band 3) determined by scanning and insertional N-glycosylation mutagenesis. Biochem. J. 339, 269–279 (1999).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the affected individuals for their cooperation; P. Christopherson for supplies of NS1652 and NS3623; M. Chetty for technical support; P. Martin for DNA sequencing; R.J. Unwin for urinary acidification data; J. Delaunay, J. Newsome, P. Williamson, P. Gover, A.B. Mehta and S. Gibbs for permission to report on individuals in their care; and D.L. Rimm for advice on software. This work was supported by the National Kidney Research Fund (G.W.S., P.H.), Action Medical Research (J.C.E.), the Wellcome Trust (J.C.E., L.J.B.), Telethon (A.I.), the UK National Health Service R & D Directorate, MURST project PRIN (Italy) and the MIUR FIRB project (A.I.). Some work was carried out at the Department of Biochemistry, University of Bristol (L.J.B.).

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Authors and Affiliations

  1. Bristol Institute for Transfusion Sciences, National Blood Service, Southmead, BS10 5ND, Bristol, UK

    Lesley J Bruce & May-Jean King

  2. University Laboratory of Physiology, South Parks Road, Oxford, OX1 3PT, UK

    Hannah C Robinson & J Clive Ellory

  3. Laboratoire de Physiologie des membranes Cellulaires, FRE 2721 CNRS-Université de Nice, Bâtiment de Sciences Naturelles, 28 av. Valrose, Nice, 06108 cedex 2, France

    Hélène Guizouarn & Franck Borgese

  4. Department of Medicine, University College London, University Street, London, WC1E 6JJ, UK

    Penny Harrison, Suzanne E Coles, Daniel M Gore & Gordon W Stewart

  5. Department of Internal Medicine, Division of Hematology, University Hospital of Zurich, Sternwartstrasse 2, Zurich, 8091, Switzerland

    Jeroen S Goede

  6. Institute of Biochemistry, ETH-Hoenggerberg, Schafmattstrasse 18, Zurich, CH-8093, Switzerland

    Hans U Lutz

  7. Telethon Institute of Genetics and Medicine, Via P. Castellino 111, Napoli, 80131, Italy

    Romina Ficarella

  8. Department of Haematology, Imperial College, Hammersmith Hospital, London, W12 0HS, UK

    D Mark Layton

  9. Department of Biochemistry and Medical Biotechnologies, CEINGE Biotecnologie Avanzante, Federico II University, Via Comunale Margherita 482, Napoli, 80131, Italy

    Achille Iolascon

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  1. Lesley J Bruce
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Corresponding author

Correspondence to Gordon W Stewart.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Schematic diagram of band 3 membrane topology. (PDF 49 kb)

Supplementary Fig. 2

Family trees. (PDF 45 kb)

Supplementary Fig. 3

Blood films. (PDF 2933 kb)

Supplementary Fig. 4

Time course of increase in plasma [K] on storage of whole heparinised blood at 37, 20 and 0°C. (PDF 115 kb)

Supplementary Table 1

Further pedigrees in which SLC4A1 was sequenced. (PDF 24 kb)

Supplementary Table 2

Ion flux and hematological data in unpublished cryohydrocytosis cases. (PDF 38 kb)

Supplementary Table 3

Ion flux and hematological data in unpublished spherocytic cases. (PDF 75 kb)

Supplementary Note

Clinical details of previously unpublished patients. (PDF 19 kb)

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Bruce, L., C Robinson, H., Guizouarn, H. et al. Monovalent cation leaks in human red cells caused by single amino-acid substitutions in the transport domain of the band 3 chloride-bicarbonate exchanger, AE1. Nat Genet 37, 1258–1263 (2005). https://doi.org/10.1038/ng1656

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