Skip to main content
Log in

Spin probe clustering in human erythrocyte ghosts

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

A model has been developed for 5-nitroxide stearate, I(12,3), distribution in human erythrocyte ghosts which accurately predicts ESR spectral alterations observed with increased probe/total lipid (P/L) at 37°C. This spin probe occupies a class of high-affinity, noninteracting sites at low loading. Saturation occurs with increasing probe concentration, and, at higher loading, the probe inserts itself at initially dilute sites to form membranebound clusters of variable size. No ‘low’ probe remains at high P/L where all I(12,3) clusters in a ‘concentrated’ phase. This model allows determination of the dilute/clustered probe ratio, and shows that I(12,3) segregates in erythrocytes at what might otherwise be considered low P/L (e.g., 1/359). These findings validate the earlier use of empirical parameters to estimate probe sequestration in biological membranes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Amatruda, J.M., Finch, F.D. 1979. Modulation of hexose uptake and insulin action by cell membrane fluidity.J. Biol. Chem. 254:2619–2625

    Google Scholar 

  2. Bieri, V., Wallach, D.F.H., Lin, P. 1974. Focal erythrocyte membrane perturbations caused by nitroxide lipid analogues.Proc. Natl. Acad. Sci. USA 71:4797–4801

    Google Scholar 

  3. Butler, K.W., Tattrie, N.H., Smith, I.C.P. 1974. The location of spin probes in two phase mixed lipid systems.Biochim. Biophys. Acta 363:351–360

    Google Scholar 

  4. Butler, K.W., Deslauriers, R., Smith, I.C.P. 1984.Plasmodium berghei: Electron spin resonance and lipid analysis of infected mouse erythrocyte membranes.Exp. Parasitol. 57:178–184

    Google Scholar 

  5. Butterfield, D.A., Whisnant, C.C., Chesnut, D.B. 1976. On the use of the spin labeling technique in the study of erythrocyte membranes.Biochim. Biophys. Acta 426:697–702

    Google Scholar 

  6. Curtain, C.C. 1984. Glycosphingolipid domain formation and lymphocyte activation.In: Biomembranes. M. Kates and L. Manson, editors. Vol. 12, pp. 603–632. Plenum, New York

    Google Scholar 

  7. Curtain, C.C., Gordon, L.M. 1984. Membrane ESR spectroscopy.In: Membranes, Detergents, and Receptor Solubilization. J.C. Venter and L. Harrison, editors. Vol. 1, pp. 177–213. Alan R. Liss, New York

    Google Scholar 

  8. Curtain, C.C., Looney, F.D., Marchalonis, J.J., Raison, J.K. 1978. Changes in lipid ordering and state of aggregation in lymphocyte plasma membranes after exposure to mitogens.J. Membrane Biol. 44:211–232

    Google Scholar 

  9. Curtain, C.C., Looney, F.D., Regan, D.L., Ivancic, N.M. 1983. Changes in the ordering of lipids in the membrane ofDunaliella in response to osmotic-pressure changes.Biochem. J. 213:131–136

    Google Scholar 

  10. Curtain, C.C., Looney, F.D., Smelstorius, J.A. 1980. Lipid domain formation and ligand-induced lymphocyte membrane changes.Biochim. Biophys. Acta 596:43–56

    Google Scholar 

  11. Curtain, C.C., Looney, F.D., Smelstorius, J.A. 1981. Glycosphingolipid clustering and mast cell degranulation.Int. Arch. Appl. Immunol. 65:34–41

    Google Scholar 

  12. Dodge, J.T., Mitchell C., Hanahan, D.J. 1963. The preparation and characterization of hemoglobin-free ghosts of human erythrocytes.Arch. Biochem. Biophys. 100:119–130

    Google Scholar 

  13. Elgavish, A., Rifkind, J., Sacktor, B. 1983. In vitro effects of vitamin D3 on the phospholipids of isolated renal brush border membranes.J. Membrane Biol. 72:85–91

    Google Scholar 

  14. Gaffney, B.J. 1974. Spin-label measurements in membranes.Methods Enzymol. 32B:161–198

    Google Scholar 

  15. Golan, D.E., Alecio, M.R., Veatch, W.R., Rando, R.R. 1984. Lateral mobility of phospholipid and cholesterol in human erythrocyte membrane: Effects of protein-lipid interactions.Biochemistry 23:332–339

    Google Scholar 

  16. Gordon, L.M. Mobley, P.W. 1984. Thermotropic lipid phase separations in human erythrocyte ghosts and cholesterol-enriched rat liver plasma membranes.J. Membrane Biol. 79:75–86

    Google Scholar 

  17. Gordon, L.M., Mobley, P.W. 1985. Membrane lipids, membrane fluidity and enzyme activity.In: Membrane Fluidity in Biology. R. C. Aloia and J. Boggs, editors. Vol. IV. Academic, New York (in press)

    Google Scholar 

  18. Gordon, L.M., Mobley, P.W., Esgate, J.A., Hofmann, G., Whetton, A.D., Houslay, M.d. 1983. Thermotropic lipid phase separations in human platelet and rat liver plasma membranes.J. Membrane Biol. 76:139–149

    Google Scholar 

  19. Gordon, L.M., Sauerheber, R.D. 1977. Studies on spin-labelled egg lecithin dispersionsBiochim. Biophys. Acta 466:34–43

    Google Scholar 

  20. Gordon, L.M., Sauerheber, R.D., Esgate, J.A. 1978. Spinlabel studies on rat liver and heart plasma membranes: Effects of temperature, calcium and lanthanum on membrane fluidity.J. Supramol. Struct. 9:299–326

    Google Scholar 

  21. Gordon, L.M., Sauerheber, R.D. Esgate, J.A., Dipple, I., Marchmont, R.J., Houslay, M.D. 1980. The increase in bilayer fluidity of rat liver plasma membranes achieved by the local anesthetic benzyl alcohol affects the activity of intrinsic membrane enzymes.J. Biol. Chem. 255:4519–4527

    Google Scholar 

  22. Griffith, O.H., Jost, P.C. 1976. Lipid spin labels in biological membranes.In: Spin Labeling. Theory and Applications. L. Berliner, editor. Chap. 7, pp. 453–523. Academic, London

    Google Scholar 

  23. Hauser, H., Guyer, W., Howell, K. 1979. Lateral distribution of negatively charged lipids in lecithin membranes. Clustering of fatty acids.Biochemistry 18:3285–3291

    Google Scholar 

  24. Horvath, L.I., Bagyinka, C.S., Sandor, M., Gergely, J. 1982 Changes in the lateral ordering of the macrophage plasma membrane during Fc receptor mediated phagocytosis.Mol. Immunol. 19:1603–1610

    Google Scholar 

  25. Houslay, M.D., Gordon, L.M. 1983. The activity of adenylate cyclase is regulated by the nature of its lipid environment.Curr. Top. Membr. Transp. 18:179–231

    Google Scholar 

  26. Houslay, M.D., Stanley, K.K. 1982. Dynamics of Biological Membranes. pp. 1–33. Wiley, New York

    Google Scholar 

  27. Humphries, G.M.K., McConnell, H.M. 1982. Nitroxide spin labels.In: Methods of Experimental Physics. R. Favia, editor. Vol. 20, pp. 53–122. Academic, New York

    Google Scholar 

  28. Koszo, F., Horvath, L.I., Simon, N., Siklosi, C.S., Kiss, M. 1982. The role of possible membrane damage in porphyrin cutanea tarda: A spin-label study of rat liver cell membranes.Biochem. Pharmacol. 31:11–17

    Google Scholar 

  29. Merritt, M.V., Loughman, B.E. 1979. Spin labeling studies of the interaction of P815 mouse mastocytoma cells with antisera to histocompatibility antigen, H-2d.Immunopharmacology 1:301–314

    Google Scholar 

  30. Pauls, K.P., Thompson, J.E., Lepock, J.R. 1980. Spin label studies of microsomal membranes fromAcanthamoeba castellanii in different states of differentiation.Arch. Biochem. Biophys. 200:22–30

    Google Scholar 

  31. Puskin, J.S., Wiese, M.B. 1982. A spin label study of human lens membranes.Exp. Eye Res. 35:251–258

    Google Scholar 

  32. Sackmann, E., Trauble, H. 1972. Studies of the crystallineliquid crystalline phase transition of lipid model membranes.J. Am. Chem. Soc. 94:4482–4491

    Google Scholar 

  33. Sauerheber, R.D., Gordon, L.M., Crosland, R.D., Kuwahara, M.D. 1977. Spin-label studies on rat liver and heart plasma membranes: Do probe-probe interactions interfere with the measurement of membrane properties?J. Membrane Biol. 31:131–169

    Google Scholar 

  34. Sauerheber, R.D., Lewis, U.J., Esgate, J.A., Gordon, L.M. 1980. Effects of calcium, insulin and growth hormone on membrane fluidity. A spin label study of rat adipocyte and human erythrocyte ghosts.Biochim. Biophys. Acta 597:292–304

    Google Scholar 

  35. Sauerheber, R.D., Zimmerman, T.S., Esgate, J.A., Vanderlaan, W.P., Gordon, L.M. 1980. Effects of calcium, lanthanum and temperature on the fluidity of spin-labeled human platelets.J. Membrane Biol. 52:201–219

    Google Scholar 

  36. Scandella, C.J., Devaux, P., McConnell, H.M. 1972. Rapid lateral diffusion of phospholipids in rabbit sarcoplasmic reticulum.Proc. Natl. Acad. Sci. USA 69:2056–2060

    Google Scholar 

  37. Stier, A., Sackmann, E. 1973. Spin labels as enzyme substrates. Heterogeneous lipid distribution in liver microsomal membranes.Biochim. Biophys. Acta 311:400–408

    Google Scholar 

  38. Thompson, N.L., Axelrod, D. 1980. Reduced lateral mobility of a fluorescent lipid probe in cholesterol-depleted erythrocyte membranes.Biochim. Biophys. Acta 597:155–165

    Google Scholar 

  39. Verma, S.P., Wallach, D.F.H. 1975. Evidence for constrained lipid mobility in the erythrocyte ghost. A spin-label study.Biochim. Biophys. Acta 382:73–82

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gordon, L.M., Looney, F.D. & Curtain, C.C. Spin probe clustering in human erythrocyte ghosts. J. Membrain Biol. 84, 81–95 (1985). https://doi.org/10.1007/BF01871650

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01871650

Key Words

Navigation