Erythrocyte membrane deformability and stability: two distinct membrane properties that are independently regulated by skeletal protein associations

Skeletal proteins play an important role in determining erythrocyte membrane biophysical properties. To study whether membrane deformability and stability are regulated by the same or different skeletal protein interactions, we measured these two properties, by means of ektacytometry, in biochemically perturbed normal membranes and in membranes from individuals with known erythrocyte abnormalities. Treatment with 2,3-diphosphoglycerate resulted in membranes with decreased deformability and decreased stability, whereas treatment with diamide produced decreased deformability but increased stability. N- ethylmaleimide induced time-dependent changes in membrane stability. Over the first minute, the stability increased; but with continued incubation, the membranes became less stable than control. Meanwhile, the deformability of these membranes decreased with no time dependence. Biophysical measurements were also carried out on pathologic erythrocytes. Membranes from an individual with hereditary spherocytosis and a defined abnormality in spectrin-protein 4.1 association showed decreased stability but normal deformability. In a family with hereditary elliptocytosis and an abnormality in spectrin self-association, the membranes had decreased deformability and stability. Finally, membranes from several individuals with Malaysian ovalocytosis had decreased deformability but increased stability. Our data from both pathologic membranes and biochemically perturbed membranes show that deformability and stability change with no fixed relationship to one another. These findings imply that different skeletal protein interactions regulate membrane deformability and stability. In light of these data, we propose a model of the role of skeletal protein interactions in deformability and stability.

[1]  S B Shohet,et al.  The influence of membrane skeleton on red cell deformability, membrane material properties, and shape. , 1983, Seminars in hematology.

[2]  W. N. Burnette,et al.  "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. , 1981, Analytical biochemistry.

[3]  N. Mohandas,et al.  Deformability of isolated red blood cell membranes. , 1982, Biochimica et biophysica acta.

[4]  D. Branton,et al.  Purification of two spectrin-binding proteins: biochemical and electron microscopic evidence for site-specific reassociation between spectrin and bands 2.1 and 4.1. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[5]  N. Mohandas,et al.  Rigid membranes of Malayan ovalocytes: a likely genetic barrier against malaria. , 1984, Blood.

[6]  G. R. Bartlett.  Phosphorus assay in column chromatography. , 1959, The Journal of biological chemistry.

[7]  N. Mohandas,et al.  Unique alpha-spectrin mutant in a kindred with common hereditary elliptocytosis. , 1987, The Journal of clinical investigation.

[8]  M. Sheetz,et al.  2,3-Diphosphoglycerate and ATP dissociate erythrocyte membrane skeletons. , 1980, The Journal of biological chemistry.

[9]  R. Josephs,et al.  Ultrastructure of unit fragments of the skeleton of the human erythrocyte membrane , 1984, The Journal of cell biology.

[10]  L. Wolfe,et al.  A genetic defect in the binding of protein 4.1 to spectrin in a kindred with hereditary spherocytosis. , 1982, The New England journal of medicine.

[11]  N. Mohandas,et al.  Analysis of factors regulating erythrocyte deformability. , 1980, The Journal of clinical investigation.

[12]  W. Groner,et al.  Ektacytometric analysis of factors regulating red cell deformability. , 1980, Blood cells.

[13]  Vann Bennett,et al.  Partial deficiency of erythrocyte spectrin in hereditary spherocytosis , 1985, Nature.

[14]  E. Evans,et al.  Adhesivity and rigidity of erythrocyte membrane in relation to wheat germ agglutinin binding , 1984, The Journal of cell biology.

[15]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[16]  V. Marchesi.  The red cell membrane skeleton: recent progress. , 1983, Blood.

[17]  J. Palek,et al.  Sulfhydryl reagents induce altered spectrin self-association, skeletal instability, and increased thermal sensitivity of red cells. , 1983, Blood.

[18]  V. Bennett,et al.  Identification and partial purification of ankyrin, the high affinity membrane attachment site for human erythrocyte spectrin. , 1979, The Journal of biological chemistry.

[19]  M. Stöhr,et al.  Selective alteration of erythrocyte deformabiliby by SH-reagents: evidence for an involvement of spectrin in membrane shear elasticity. , 1978, Biochimica et biophysica acta.

[20]  C. Haest,et al.  Spectrin as a stabilizer of the phospholipid asymmetry in the human erythrocyte membrane. , 1978, Biochimica et biophysica acta.

[21]  P. L. la Celle,et al.  Abnormalities in the membrane material properties of hereditary spherocytes. , 1980, Journal of biomechanical engineering.

[22]  N. Mohandas,et al.  A technique to detect reduced mechanical stability of red cell membranes: relevance to elliptocytic disorders. , 1982, Blood.

[23]  C. M. Cohen.  The molecular organization of the red cell membrane skeleton. , 1983, Seminars in hematology.

[24]  D. Branton,et al.  Interaction of cytoskeletal proteins on the human erythrocyte membrane , 1981, Cell.

[25]  J. Prchal,et al.  Altered spectrin dimer-dimer association and instability of erythrocyte membrane skeletons in hereditary pyropoikilocytosis. , 1981, The Journal of clinical investigation.

[26]  R. Waugh.  Temperature dependence of the yield shear resultant and the plastic viscosity coefficient of erythrocyte membrane. Implications about molecular events during membrane failure. , 1982, Biophysical journal.

[27]  S B Shohet,et al.  Deficiency of skeletal membrane protein band 4.1 in homozygous hereditary elliptocytosis. Implications for erythrocyte membrane stability. , 1981, The Journal of clinical investigation.

[28]  Shih-Chun Liu,et al.  Spectrin tetramer–dimer equilibrium and the stability of erythrocyte membrane skeletons , 1980, Nature.

[29]  D. Speicher,et al.  Identification of functional domains of human erythrocyte spectrin. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[30]  B ZAK,et al.  A new method for the direct determination of serum cholesterol. , 1953, The Journal of laboratory and clinical medicine.

[31]  N. Mohandas,et al.  Erythrocyte membrane rigidity induced by glycophorin A-ligand interaction. Evidence for a ligand-induced association between glycophorin A and skeletal proteins. , 1985, The Journal of clinical investigation.