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. 1982 Feb 1;201(2):259–266. doi: 10.1042/bj2010259

Hereditary spherocytosis of man. Altered binding of cytoskeletal components to the erythrocyte membrane

John S Hill *, William H Sawyer *, Geoffrey J Howlett *, James S Wiley †
PMCID: PMC1163639  PMID: 7082289

Abstract

Human erythrocytes possess a lattice work of extrinsic proteins on the inner face of the membrane (`cytoskeleton') that maintains the shape and deformability of the cell. The major proteins of the cytoskeleton are spectrin and actin, which are attached to the membrane by protein bands 2.1 (`ankyrin') and 4.1. The interactions of spectrin/actin with erythrocyte membranes from normal subjects and from patients with hereditary spherocytosis (HS) have been studied by using an air-driven ultracentrifuge, which can rapidly separate membranes from soluble proteins (150000g for 30s). The total amount of spectrin/actin in HS and normal ghosts is similar. However, the rate of dissociation of spectrin and actin from HS erythrocyte membranes at low ionic strength is significantly lower than that observed for normal membranes. Spectrin and actin isolated from either HS or normal membranes re-associated in a similar manner to spectrin/actin-depleted vesicles prepared from normal cells. Scatchard analysis showed an average binding capacity of 278μg/mg of membrane protein. However, spectrin/actin-depleted vesicles prepared from HS cells bound significantly less spectrin/actin prepared from either the normal or abnormal cells (average binding capacity 158μg/mg of membrane protein). The defect was defined further by studying the cytoskeleton obtained by Triton X-100 extraction of membranes. Under conditions of low ionic strength cytoskeletons prepared from HS membranes dissociated more slowly than those prepared from normal membranes, and only 80% of the protein from HS cytoskeletons could be solubilized after 180min compared with 100% for normal cytoskeletons. The difference between HS and normal membranes, which persists in isolated cytoskeletons, suggests that alterations in either the primary structure or the degree of phosphorylation of protein bands 2.1 or 4.1 may be central to the molecular basis of hereditary spherocytosis.

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Selected References

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

  1. Anderson J. M., Tyler J. M. State of spectrin phosphorylation does not affect erythrocyte shape or spectrin binding to erythrocyte membranes. J Biol Chem. 1980 Feb 25;255(4):1259–1265. [PubMed] [Google Scholar]
  2. BERTLES J. F. Sodium transport across the surface membrane of red blood cells in hereditary spherocytosis. J Clin Invest. 1957 Jun;36(6 Pt 1):816–824. doi: 10.1172/JCI103487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett V., Branton D. Selective association of spectrin with the cytoplasmic surface of human erythrocyte plasma membranes. Quantitative determination with purified (32P)spectrin. J Biol Chem. 1977 Apr 25;252(8):2753–2763. [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Cooper R. A., Jandl J. H. The role of membrane lipids in the survival of red cells in hereditary spherocytosis. J Clin Invest. 1969 Apr;48(4):736–744. doi: 10.1172/JCI106031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cooper R. A., Jandl J. H. The selective and conjoint loss of red cell lipids. J Clin Invest. 1969 May;48(5):906–914. doi: 10.1172/JCI106049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cooper R. A., Sawyer W. H., Leslie M. H., Hill J. S., Gill F. M., Wiley J. S. Normal fluidity of red cell membranes in hereditary spherocytosis. Br J Haematol. 1980 Oct;46(2):299–301. doi: 10.1111/j.1365-2141.1980.tb05969.x. [DOI] [PubMed] [Google Scholar]
  8. DE GIER J., VAN DEENENL, VERLOOP M. C., VAN GASTEL PHOSPHOLIPID AND FATTY ACID CHARACTERISTICS OF ERYTHROCYTES IN SOME CASES OF ANAEMIA. Br J Haematol. 1964 Apr;10:246–256. doi: 10.1111/j.1365-2141.1964.tb00699.x. [DOI] [PubMed] [Google Scholar]
  9. ELLMAN G. L., COURTNEY K. D., ANDRES V., Jr, FEATHER-STONE R. M. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961 Jul;7:88–95. doi: 10.1016/0006-2952(61)90145-9. [DOI] [PubMed] [Google Scholar]
  10. Golan D. E., Veatch W. Lateral mobility of band 3 in the human erythrocyte membrane studied by fluorescence photobleaching recovery: evidence for control by cytoskeletal interactions. Proc Natl Acad Sci U S A. 1980 May;77(5):2537–2541. doi: 10.1073/pnas.77.5.2537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Greenquist A. C., Shohet S. B. Phosphorylation in erythrocyte membranes from abnormally shaped cells. Blood. 1976 Dec;48(6):877–886. [PubMed] [Google Scholar]
  12. Hargreaves W. R., Giedd K. N., Verkleij A., Branton D. Reassociation of ankyrin with band 3 in erythrocyte membranes and in lipid vesicles. J Biol Chem. 1980 Dec 25;255(24):11965–11972. [PubMed] [Google Scholar]
  13. JACOB H. S., JANDL J. H. INCREASED CELL MEMBRANE PERMEABILITY IN THE PATHOGENESIS OF HEREDITARY SPHEROCYTOSIS. J Clin Invest. 1964 Aug;43:1704–1720. doi: 10.1172/JCI105046. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kuiper P. J., Livne A. Differences in fatty acid composition between normal human erythrocytes and hereditary spherocytosis affected cells. Biochim Biophys Acta. 1972 Apr 18;260(4):755–758. doi: 10.1016/0005-2760(72)90027-6. [DOI] [PubMed] [Google Scholar]
  15. LaCelle P. L. Alteration of membrane deformability in hemolytic anemias. Semin Hematol. 1970 Oct;7(4):355–371. [PubMed] [Google Scholar]
  16. Lange Y., Cutler H. B., Steck T. L. The effect of cholesterol and other intercalated amphipaths on the contour and stability of the isolated red cell membrane. J Biol Chem. 1980 Oct 10;255(19):9331–9337. [PubMed] [Google Scholar]
  17. Lux S. E. Dissecting the red cell membrane skeleton. Nature. 1979 Oct 11;281(5731):426–429. doi: 10.1038/281426a0. [DOI] [PubMed] [Google Scholar]
  18. Matsumoto N., Yawata Y., Jacob H. S. Association of decreased membrane protein phosphorylation with red blood cell spherocytosis. Blood. 1977 Feb;49(2):233–239. [PubMed] [Google Scholar]
  19. Murphy J. R. The influence of pH and temperature on some physical properties of normal erythrocytes and erythrocytes from patients with hereditary spherocytosis. J Lab Clin Med. 1967 May;69(5):758–775. [PubMed] [Google Scholar]
  20. Palek J., Liu S. C. Dependence of spectrin organization in red blood cell membranes on cell metabolism: implications for control of red cell shape, deformability, and surface area. Semin Hematol. 1979 Jan;16(1):75–93. [PubMed] [Google Scholar]
  21. Peacock A. C., Dingman C. W. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry. 1968 Feb;7(2):668–674. doi: 10.1021/bi00842a023. [DOI] [PubMed] [Google Scholar]
  22. Plut D. A., Hosey M. M., Tao M. Evidence for the participation of cytosolic protein kinases in membrane phosphorylation in intact erythrocytes. Eur J Biochem. 1978 Jan 16;82(2):333–337. doi: 10.1111/j.1432-1033.1978.tb12027.x. [DOI] [PubMed] [Google Scholar]
  23. ROBERTSON A. F., LANDS W. E. METABOLISM OF PHOSPHOLIPIDS IN NORMAL AND SPHEROCYTIC HUMAN ERYTHROCYTES. J Lipid Res. 1964 Jan;5:88–93. [PubMed] [Google Scholar]
  24. Reed C. F. Incorporation of orthophosphate-32P into erythrocyte phospholipids in normal subjects and in patients with hereditary spherocytosis. J Clin Invest. 1968 Dec;47(12):2630–2638. doi: 10.1172/JCI105946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robinson J., McCann S., McKillen M. N. Human erythrocyte hereditary spherocytosis: a membrane defect? The continuing enigma [proceedings]. Biochem Soc Trans. 1979 Jun;7(3):531–533. doi: 10.1042/bst0070531. [DOI] [PubMed] [Google Scholar]
  26. Schrier S. L., Ben-Bassat I., Bensch K., Seeger M., Junga I. Erythrocyte membrane vacuole formation in hereditary spherocytosis. Br J Haematol. 1974 Jan;26(1):59–69. doi: 10.1111/j.1365-2141.1974.tb00449.x. [DOI] [PubMed] [Google Scholar]
  27. Sheehy R., Ralston G. B. Abnormal binding of spectrin to the membrane of erythrocytes in some cases of hereditary spherocytosis. Blut. 1978 Mar 15;36(3):145–148. doi: 10.1007/BF00996653. [DOI] [PubMed] [Google Scholar]
  28. Sheetz M. P., Sawyer D. Triton shells of intact erythrocytes. J Supramol Struct. 1978;8(4):399–412. doi: 10.1002/jss.400080403. [DOI] [PubMed] [Google Scholar]
  29. Shohet S. B. Reconstitution of spectrin-deficient, spherocytic mouse erythrocyte membranes. J Clin Invest. 1979 Aug;64(2):483–494. doi: 10.1172/JCI109486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tange T., Chiba S. Rough endoplasmic reticulum-associated tubular structures occurring in acute promyelocytic leukaemia. Scand J Haematol. 1979 Jul;23(1):25–29. doi: 10.1111/j.1600-0609.1979.tb02848.x. [DOI] [PubMed] [Google Scholar]
  31. Tyler J. M., Reinhardt B. N., Branton D. Associations of erythrocyte membrane proteins. Binding of purified bands 2.1 and 4.1 to spectrin. J Biol Chem. 1980 Jul 25;255(14):7034–7039. [PubMed] [Google Scholar]
  32. Ungewickell E., Bennett P. M., Calvert R., Ohanian V., Gratzer W. B. In vitro formation of a complex between cytoskeletal proteins of the human erythrocyte. Nature. 1979 Aug 30;280(5725):811–814. doi: 10.1038/280811a0. [DOI] [PubMed] [Google Scholar]
  33. Wiley J. S. Co-ordinated increase of sodium leak and sodium pump in hereditary spherocytosis. Br J Haematol. 1972 May;22(5):529–542. doi: 10.1111/j.1365-2141.1972.tb05700.x. [DOI] [PubMed] [Google Scholar]
  34. Wolfe L. C., Lux S. E. Membrane protein phosphorylation of intact normal and hereditary spherocytic erythrocytes. J Biol Chem. 1978 May 10;253(9):3336–3342. [PubMed] [Google Scholar]
  35. Yu J., Fischman D. A., Steck T. L. Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. J Supramol Struct. 1973;1(3):233–248. doi: 10.1002/jss.400010308. [DOI] [PubMed] [Google Scholar]
  36. Zail S. S., Pickering A. Fatty acid composition of erythrocytes in hereditary spherocytosis. Br J Haematol. 1979 Jul;42(3):399–402. doi: 10.1111/j.1365-2141.1979.tb01148.x. [DOI] [PubMed] [Google Scholar]
  37. Zail S. S. The erythrocyte membrane abnormality of hereditary spherocytosis. Br J Haematol. 1977 Nov;37(3):305–310. doi: 10.1111/j.1365-2141.1977.tb01000.x. [DOI] [PubMed] [Google Scholar]

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