Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1977 Jun 1;73(3):638–646. doi: 10.1083/jcb.73.3.638

On the mechanism of ATP-induced shape changes in human erythrocyte membranes. I. The role of the spectrin complex

PMCID: PMC2111425  PMID: 873993

Abstract

Human erythrocyte ghosts have been shown, by scanning electron microscopy, to undergo ATP-dependent shape changes. Under appropriate conditions the ghosts prepared from normal disk-shaped intact cells adopt a highly crenated shape, which in the presence of Mg-ATP at 37 degrees C is slowly converted to the disk shape and eventually to the cup shape. These changes are not observed with other nucleotides or with 5'-adenylyl imidodiphosphate. Anti-spectrin antibodies, incorporated along with the Mg-ATP into the ghosts in amounts less than equivalent to the spectrin, markedly accelerate the shape changes observed with the Mg-ATP alone. The Fab fragments of these antibodies, however, have no effect. The conclusion is that the structural effect produced by the ATP is promoted by the cross-linking of spectrin by its antibodies, and may therefore itself be some kind of polymerization or network formation involving the spectrin complex on the cytoplasmic face of the membrane. The factors that contribute to the shape of the ghost and of the intact erythrocyte are discussed in the light of these findings.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Avruch J., Fairbanks G. Phosphorylation of endogenous substrates by erythrocyte membrane protein kinases. I. A monovalent cation-stimulated reaction. Biochemistry. 1974 Dec 31;13(27):5507–5514. doi: 10.1021/bi00724a009. [DOI] [PubMed] [Google Scholar]
  2. Birchmeier W., Singer S. J. On the mechanism of ATP-induced shape changes in human erythrocyte membranes. II. The role of ATP. J Cell Biol. 1977 Jun;73(3):647–659. doi: 10.1083/jcb.73.3.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Canham P. B. The minimum energy of bending as a possible explanation of the biconcave shape of the human red blood cell. J Theor Biol. 1970 Jan;26(1):61–81. doi: 10.1016/s0022-5193(70)80032-7. [DOI] [PubMed] [Google Scholar]
  4. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  5. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  6. HOFFMAN J. F. Physiological characteristics of human red blood cell ghosts. J Gen Physiol. 1958 Sep 20;42(1):9–28. doi: 10.1085/jgp.42.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Loor F., Forni L., Pernis B. The dynamic state of the lymphocyte membrane. Factors affecting the distribution and turnover of surface immunoglobulins. Eur J Immunol. 1972 Jun;2(3):203–212. doi: 10.1002/eji.1830020304. [DOI] [PubMed] [Google Scholar]
  8. Nicolson G. L., Marchesi V. T., Singer S. J. The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies. J Cell Biol. 1971 Oct;51(1):265–272. doi: 10.1083/jcb.51.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nicolson G. L., Painter R. G. Anionic sites of human erythrocyte membranes. II. Antispectrin-induced transmembrane aggregation of the binding sites for positively charged colloidal particles. J Cell Biol. 1973 Nov;59(2 Pt 1):395–406. doi: 10.1083/jcb.59.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. PORTER R. R. The hydrolysis of rabbit y-globulin and antibodies with crystalline papain. Biochem J. 1959 Sep;73:119–126. doi: 10.1042/bj0730119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Palek J., Stewart G., Lionetti F. J. The dependence of shape of human erythrocyte ghosts on calcium, magnesium, and adenosine triphosphate. Blood. 1974 Oct;44(4):583–597. [PubMed] [Google Scholar]
  12. Penniston J. T., Green D. E. The conformational basis of energy transformations in membrane systems. IV. Energized states and pinocytosis in erythrocyte ghosts. Arch Biochem Biophys. 1968 Nov;128(2):339–350. doi: 10.1016/0003-9861(68)90040-4. [DOI] [PubMed] [Google Scholar]
  13. Pinder J. C., Bray D., Gratzer W. B. Actin polymerisation induced by spectrin. Nature. 1975 Dec 25;258(5537):765–766. doi: 10.1038/258765a0. [DOI] [PubMed] [Google Scholar]
  14. Pollard T. D., Weihing R. R. Actin and myosin and cell movement. CRC Crit Rev Biochem. 1974 Jan;2(1):1–65. doi: 10.3109/10409237409105443. [DOI] [PubMed] [Google Scholar]
  15. Renooij W., Van Golde L. M., Zwaal R. F., Roelofsen B., Van Deenen L. L. Preferential incorporation of fatty acids at the inside of human erythrocyte membranes. Biochim Biophys Acta. 1974 Sep 6;363(2):287–292. doi: 10.1016/0005-2736(74)90069-8. [DOI] [PubMed] [Google Scholar]
  16. Sheetz M. P., Painter R. G., Singer S. J. Biological membranes as bilayer couples. III. Compensatory shape changes induced in membranes. J Cell Biol. 1976 Jul;70(1):193–203. doi: 10.1083/jcb.70.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sheetz M. P., Painter R. G., Singer S. J. Relationships of the spectrin complex of human erythrocyte membranes to the actomyosins of muscle cells. Biochemistry. 1976 Oct 5;15(20):4486–4492. doi: 10.1021/bi00665a024. [DOI] [PubMed] [Google Scholar]
  18. Sheetz M. P., Singer S. J. Biological membranes as bilayer couples. A molecular mechanism of drug-erythrocyte interactions. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4457–4461. doi: 10.1073/pnas.71.11.4457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sheetz M. P., Singer S. J. Equilibrium and kinetic effects of drugs on the shapes of human erythrocytes. J Cell Biol. 1976 Jul;70(1):247–251. doi: 10.1083/jcb.70.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shohet S. B., Nathan D. G., Karnovsky M. L. Stages in the incorporation of fatty acids into red blood cells. J Clin Invest. 1968 May;47(5):1096–1108. doi: 10.1172/JCI105799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Singer S. J. Molecular biology of cellular membranes with applications to immunology. Adv Immunol. 1974;19(0):1–66. doi: 10.1016/s0065-2776(08)60251-5. [DOI] [PubMed] [Google Scholar]
  22. Tilney L. G., Detmers P. Actin in erythrocyte ghosts and its association with spectrin. Evidence for a nonfilamentous form of these two molecules in situ. J Cell Biol. 1975 Sep;66(3):508–520. doi: 10.1083/jcb.66.3.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weed R. I., LaCelle P. L., Merrill E. W. Metabolic dependence of red cell deformability. J Clin Invest. 1969 May;48(5):795–809. doi: 10.1172/JCI106038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Williams R. O. The phosphorylation and isolation of two erythrocyte membrane proteins in vitro. Biochem Biophys Res Commun. 1972 May 26;47(4):671–678. doi: 10.1016/0006-291x(72)90544-x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES