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Hemoglobin-depleted human erythrocyte ghosts: Characterization of morphology and transport functions

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Summary

A method of preparing hemoglobin-depleted resealed ghosts with an extremely low hemoglobin content is described. The membrane morphology, the crossed immunoelectrophoresis pattern of the membrane proteins, and the transport function of these ghosts have been examined.

Electron microscopic examination of the ghosts on hydrophilic as well as hydrophobic grid surfaces revealed a faint filamentous network (spectrin) associated with the membrane. The ghosts were found to have permeabilities towards small polar molecules (water and mannitol) and ions (chloride, sodium, and potassium) which are quantitatively very close to those of intact red cells.

It is concluded that white ghosts prepared by the present method are well suited for simultaneous studies of morphology, membrane biochemistry, and membrane transport functions.

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References

  • Bjerrum, O.J., Bøg-Hansen, T.C. 1976. The immunochemical approach to the characterization of membrane proteins.Biochim. Biophys. Acta 455:66

    Google Scholar 

  • Bjerrum, O.J., Lundahl, P. 1974. Crossed immunoelectrophoresis of human erythrocyte membrane proteins.Biochim. Biophys. Acta 342:69

    Google Scholar 

  • Bjerrum, P.J., Tranum-Jensen, J. 1977. Erythrocyte membrane, spectrin morphology and membrane permeability at normal and low pH values. Proceedings of the International Union of Physiological Science. Vol. XIII p. 80. (Paris, 1977)

    Google Scholar 

  • Bodemann, H., Passow, H. 1972. Factors controlling the reaseling of the membrane of human erythrocyte ghosts after hypotonic hemolysis.J. Membrane Biol. 8:1

    Google Scholar 

  • Bowmann, R.J., Levitt, D.G. 1977. Polyol permeability of the human red cell: Interpretation of glucose transport in terms of a pore.Biochim. Biophys. Acta 466:68

    Google Scholar 

  • Brahm, J. 1977. Temperature-dependent changes of chloride transport in human red cells.J. Gen. Physiol. 70:283

    Google Scholar 

  • Brecher, G., Jacobek, E.F., Schneiderman, M.A. 1962. Size distribution of erythrocytes. Ann. N.Y. Acad. Sci.99:242

    Google Scholar 

  • Cabantchik, Z.I., Rothstein, A. 1974. Membrane proteins related to anion permeability of human red cell.J. Membrane Biol. 15:207

    Google Scholar 

  • Cass, A., Finkelstein, A. 1967. Water permeability of thin lipid membranes.J. Gen. Physiol. 50:1765

    Google Scholar 

  • Dalmark, M., Wieth, J.O. 1972. Temperature dependence of chloride, bromide, iodide, thiocyanate and salicylate transport in human red cells.J. Physiol. (London) 224:583

    Google Scholar 

  • Dodge, J.T., Mitchell, C., Hanahan, D.J. 1963. The preparation and chemical characterization or hemoglobin-free ghosts of human erythrocytes.Arch. Biochim. Biophys. 100:119

    Google Scholar 

  • Funder, J., Tosteson, D.C., Wieth, J.O. 1978. Effects of bicarbonate on lithium transport in human red cellsJ. Gen. Physiol. 71:721

    Google Scholar 

  • Funder, J., Wieth, J.O. 1966. Determination of sodium, potassium and water in human red blood cells.Scand. J. Clin. Lab. Invest. 18:151

    Google Scholar 

  • Funder, J., Wieth, J.O. 1967. Effects of ouabain on glucose metabolism and on fluxes of sodium and potassium of human blood cells.Acta Physiol. Scand. 71:113

    Google Scholar 

  • Funder, J., Wieth, J.O. 1976. Chloride transport in human erythrocytes and ghosts: A quantitative comparison.J. Physiol. (London) 262:679

    Google Scholar 

  • Gardos, G. 1958. The function of calcium in the potassium permeability of human erythrocytes.Biochim. Biophys. Acta 30:653

    Google Scholar 

  • Gunn, R.B., Dalmark, M., Tosteson, D.C., Wieth, J.O. 1973. Characteristics of chloride transport in human red blood cells.J. Gen. Physiol. 61:185

    Google Scholar 

  • Gunn, R.B., Wieth, J.O., Tosteson, D.C. 1975. Some effects of low pH on chloride exchange in human red blood cells.J. Gen. Physiol. 65:731

    Google Scholar 

  • Hainfield, J.F. and Steck, T.L. 1977. The sub-membrane reticulum of the human erythrocyte: A scanning electron microscope study.J. Supramol. Struct. 6:301

    Google Scholar 

  • Hazlewood, C.F., Beall, P.T., Singer, D. 1978. Electron microscopic examinations of human red blood cell “ghost” preparations.Fed. Proc. 37:313

    Google Scholar 

  • Helenius, A., Simons, K. 1975. Solubilization of membranes by detergents.Biochim. Biophys. Acta 415:29

    Google Scholar 

  • Hillier, J., Hoffman, J.F. 1953. On the ultrastructure of the plasma membrane as determined by the electron microscope.J. Cell Comp. Physiol. 42:203

    Google Scholar 

  • Hoogeveen, J.T., Juliano, R., Coleman, J., Rothstein, A. 1970. Water-soluble proteins of the human red cell membrane.J. Membrane Biol. 3:156

    Google Scholar 

  • Johnson, R.M. 1975. The kinetics of resealing of washed erythrocyte ghosts.J. Membrane Biol. 22:231

    Google Scholar 

  • Jung, C.Y. 1971. Evidence of high stability of the glucose transport carrier function in human red cell ghosts extensively washed in various media.Arch. Biochem. Biophys. 146:215

    Google Scholar 

  • Lepke, S., Fasold, H., Pring, M., Passow, H. 1976. A study of the relationship between inhibition of anion exchange and binding to red blood cell membrane of 4.4′-diisothiocyano stilbene-2.2′-disulfonic acid (DIDS) and its dihydro derivative (H2DIDS).J. Membrane Biol. 29:147

    Google Scholar 

  • Macey, R.I., Karan, D.M., Farmer, R.E.L. 1972. Properties of water channels in human red cells.In: Biomembranes. Vol. 3, pp. 331–340. F. Kreuzer and J.F.G. Slegers, editors. Plenum Press, New York

    Google Scholar 

  • Norrild, B., Bjerrum, O.J., Vestergaard, B.F. 1977. Polypeptide analysis of individual immunoprecipitates from crossed immunoelectrophoresis.Anal. Biochem. 81:432

    Google Scholar 

  • Rostgaard, J., Christensen, P. 1975. A multipurpose specimen-carrier for handling small biological objects through critical point drying.J. Microsc. 105:107

    Google Scholar 

  • Schwoch, G., Passow, H. 1973. Preparation and properties of human erythrocyte ghosts.Molec. Cell. Biochem. 2:197

    Google Scholar 

  • Ship, S., Shami, Y., Breuer, W., Rothstein, A. 1977. Synthesis of tritiated 4,4′-diisothiocyano-2.2′ stilbene disulfonic acid ([3H]DIDS) and its covalent reaction with sites related to anion transport in human red blood cells.J. Membrane Biol. 33:311

    Google Scholar 

  • Steck, T.L. 1974. The organization of proteins in the human red blood cell membrane.J. Cell Biol. 62:1

    Google Scholar 

  • Weed, R.I., Reed, C.F., Berg, G. 1963. Is hemoglobin an essential structural component of human erythrocyte membranes.J. Clin. Invest. 42:581

    Google Scholar 

  • Weeke, B. 1973. General remarks on principles equipment, reagents and procedures.Scand. J. Immunol. (2 suppl.)1:15

    Google Scholar 

  • Westerman, M.P., Pierce, L.E., Jensen, W.N. 1961. A direct method for the quantitative measurement of red cell dimensions.J. Lab. Clin. Med. 57:819

    Google Scholar 

  • Zaki, L., Fasold, H., Schuhmann, B., Passow, H. 1975. Chemical Modification of membrane proteins in relation to inhibition of anion exchange in human red blood cells.J. Cell. Physiol. 86:471

    Google Scholar 

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Bjerrum, P.J. Hemoglobin-depleted human erythrocyte ghosts: Characterization of morphology and transport functions. J. Membrain Biol. 48, 43–67 (1979). https://doi.org/10.1007/BF01869256

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  • DOI: https://doi.org/10.1007/BF01869256

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