Summary
We have sought to elucidate the spiculated shape of McLeod erythrocytes. Red cells from a normal donor and from a McLeod patient were incubated in phosphate-buffered saline containing 0, 0.05, or 0.1mm chlorpromazine at 0°C for 5 min. then glutaraldehyde-fixed, and examined by scanning electron microscopy. The normal red cells were biconcave disks in which chlorpromazine induced inward (negative) curvature: deep cupping (stomatocytosis) and multiple invaginations. The McLeod cells were mostly spiculated. Chlorpromazine at lower concentration converted them into biconcave disks and, at higher concentration, into stomatocytes. These results support the hypothesis that the spiculation of McLeod cells is the result of an imbalance of surface area between the two lipid leaflets of the membrane; that is, a bilayer couple effect.
We determined the numerical density of intramembrane particles (IMP) in replicas of both fracture faces of red cells subjected to freeze fracture and rotary shadowing. These values were as follows (expressed per μm2 of membrane ±sd): the normal protoplasmic fracture face had 2200±306 and the McLeod had 2300±250. The normal exoplasmic fracture face had 388±75 and the McLeod had 330±59. We conclude that there is no evidence for derangement of band 3, the principal protein in theIMP, in McLeod erythrocytes.
Similar content being viewed by others
References
Agré, P., Casella, J.F., Zinkham, W.H., McMillan, C., Bennett, V. 1985. Partial deficiency of erythrocyte spectrin in hereditary spherocytosis.Nature (London) 314:380–383
Agré, P., Orringer, E.P., Chui, D.H.K., Bennett, V. 1981. A molecular defect in two families with hemolytic poikilocytic anemia. Reduction of high affinity membrane binding sites for ankyrin.J. Clin. Invest. 68:1566–1576
Allen, F.H., Krabbe, S.M.R., Corcoran, P.A. 1961. A new phenotype (McLeod) in the Kell blood group system.Vox Sang. 6:555–560
Bessis, M. 1973.In: Red Cell Shape. M. Bessis, R.I. Weed, and P.F. LeBlond, editors. pp. 1–23. Springer-Verlag, New York
Branton, D., Cohen, C.M., Tyler, J. 1981. Interaction of cytoskeletal proteins on the human erythrocyte membrane.Cell 24:24–32
Chaplin, D.B., Kleinfeld, A.M. 1983. Interaction of fluorescent quenchers with the n-(9-anthroyloxyl) fatty acid membrane probes.Biochim. Biophys. Acta 73:465–474
Conrad, M.J., Singer, S.J. 1981. The solubility of amphipathic molecules in biological membranes and lipid bilayers and its implications for membrane structure.Biochemistry 20:808–818
Danon, D., Goldstein, L., Marikovsky, Y., Skuletsky, E. 1972. Use of cationized ferritin as a label of negative charges on cell surfaces.J. Ultrastruct. Res. 38:500–510
Ferrell, J.E., Jr., Lee, K.J., Huestis, W.H. 1985. Membrane bilayer balance and erythrocyte shape: A quantitative assessment.Biochemistry 24:2849–2857
Galey, W.R., Evan, A.P., Van Nice, P.S., Dail, W.G., Cooper, R.A. 1978. Morphology and physiology of the McLeod erythrocyte: I. Scanning electron microscopy and water transport properties.Vox Sang. 34:152–161
Glaubensklee, C.S., Evan, A.P., Galey, R. 1982. Structural and biochemical analysis of the McLeod erythrocyte membrane.Vox Sang. 42:262–271
Jinbu, Y., Sato, S., Nakao, T., Nakao, M. 1982. Ankyrin is necessary for both drug-induced and ATP-induced shape change of human erythrocyte ghosts.Biochem. Biophys. Res. Commun. 104:1087–1092
Khodadad, J.K., Loew, J.M., Weinstein, R.S. 1986. Freezefracture and freeze-etch electron microscopy of membrane proteins.In: Techniques for the Analysis of Membrane Proteins. C. I. Ragan and R. J. Cherry, editors. pp. 275–314. Chapman and Hall, London
Khodadad, J.K., Weinstein, R.S. 1983. The band 3-rich membrane of llama erythrocyte: Studies on cell shape and the organization of membrane proteins.J. Membrane Biol. 72:161–171
Khodadad, J.K., Weinstein, R.S., Steck, T.L. 1986. Quantitation of intramembrane particles of McLeod red cells.Proc. 44th Annu. Meet. Elect. Microsc. Soc. Am. (Albuquerque, NM) pp. 218–219
Kuypers, F.A., Linde-Sibenius Trip, M., Roelofsen, B., Opden Kamp, J.A.F., Tanner, M.J.A., Anstee, D.J. 1985. The phospholipid organization in the membranes of McLeod and Leach phenotype erythrocytes.FEBS Lett. 184:20–24
Lange, Y., Steck, T.L. 1984. Mechanism of red blood cell acanthocytosis and echinocytosis in vivo.J. Membrane Biol. 77:153–159
Lawler, J., Liu, S.C., Palek, J., Prchal, J. 1984. A molecular defect of spectrin in a subset of patients with hereditary elliptocytosis. Alterations in the a-subunit domain involved in spectrin self association.J. Clin. Invest. 73:1688–1984
Lieber, M.R., Lange, Y., Weinstein, R.S., Steck, T.L. 1984. Interaction of chlorpromazine with the human erythrocyte membrane.J. Biol. Chem. 259:9225–9234
Lux, S.E. 1979. Spectrin-actin membrane skeleton of normal and abnormal red blood cells.Semin. Hematol. 16:21–51
Luxnat, M., Müller, H.J., Galla, H.-J. 1984. Membrane solubility of chlorpromazine.Biochem. J. 224:1023–1026
Marchesi, S.L., Knowles, W.J., Morrow, J.S., Bologna, M., Marchesi, V.T. 1986. Abnormal spectrin in hereditary elliptocytosis.Blood 67:141–151
Margaritis, L.H., Elgsaeter, A., Branton, D. 1977. Rotary replication for freeze-etching.J. Cell Biol. 72:47–56
Marsh, W.L., Oyen, R., Nichols, M.E., Allen, F.H., Jr., 1975. Chronic granulomatous disease and the Kell blood groups.Br. J. Haematol. 29:247–262
Marsh, W.L., Redman, C.M. 1987. Recent developments in the Kell blood group system.Transfus. Med. Rev. 1:4–20
Moor, H., Mühlethaler, K. 1963. Fine structure in frozen etched yeast cells.J. Cell Biol. 17:609–628
Parker, J.D., Burkowitz, L.R. 1986. Genetic variants affecting the structure and function of human red cell membrane.In: Physiology of Membrane Disorders. (2nd ED.) T.E. Andreoli, J.F. Hoffman, D.D. Fanestil, and S.G. Shultz, editors. pp. 785–814. Plenum, New York-London
Pjura, W.J., Kleinfeld, A.M., Karnovsky, M.J. 1984. Partition of fatty acids and fluorescent fatty acids into membranes.Biochemistry 23:2039–2043
Redman, C.M., Marsh, W.L., Scarborough, A., Johnson, C.L., Rabin, B.I., Overbeeke, M. 1988. Biochemical studies on McLeod phenotype red cells and isolation of Kx Antigen.Br. J. Haematol. 68:131–136
Sheetz, M.P., Singer, S.J. 1974. Biological membranes as bilayer couples. A molecular mechanism of drug-erythrocyte interaction.Proc. Natl. Acad. Sci. USA 71:4457–4461
Singer, K., Fisher, B., Perlstein, M.A. 1952. Acanthocytosis: Genetic erythrocytic malformation.Blood 7:577–591
Steck, T.L. 1989. Red cell shape.In: Cell Shape: Determinants, Regulation, Regulatory Role. W. Stein and F. Bonner, editors. Academic, New York (in press)
Tang, L.L., Redman, C.M., Williams, D., Marsh, W.L. 1981. Biochemical studies on McLeod phenotype erythrocytes.Vox Sang. 40:17–26
Weinstein, R.S., Benefiel, D.J., Pauli, B.U. 1979. Use of computers in the analysis of intramembrane particles.In: Freeze Fracture: Methods, Artifacts and Interpretations. J.E. Rash and C.S. Hudson, editors. pp. 175–183. Raven, New York
Weinstein, R.S., Khodadad, J.K., Steck, T.L. 1978. Fine structure of the band 3 protein in human red cell membrane. Freeze-fracture studies.J. Supramol. Struct. 8:325–335
Weinstein, R.S., Khodadad, J.K., Steck, T.L. 1978. Ultrastructural characterization of proteins at the natural surfaces of the red cell membrane.Prog. Clin. Biol. Res. 21:413–427
Weinstein, R.S., Khodadad, J.K., Steck, T.L. 1980. The band 3 protein particle of the human red blood cell.In: Membrane Transport in Erythrocytes. U.V. Lassen, H.H. Ussing, and J.O. Wieth, editors. pp. 35–50. Munksgaard, Copenhagen
Wimer, B.M., Marsh, W.L., Taswell, H.F., Galey, W.R. 1977. Haematological changes associated with the McLeod phenotype of the Kell blood group system.Br. J. Haematol. 36:219–224
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Khodadad, J.K., Weinstein, R.S., Marsh, L.W. et al. Shape determinants of McLeod acanthocytes. J. Membrain Biol. 107, 213–218 (1989). https://doi.org/10.1007/BF01871936
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01871936