The Journal of Membrane Biology

, Volume 87, Issue 1, pp 35–44 | Cite as

Melittin lysis of red cells

  • M. T. Tosteson
  • S. J. Holmes
  • M. Razin
  • D. C. Tosteson


This paper describes experiments designed to explore interactions between human red blood cell membranes and melittin, the main component of bee venom. We found that melittin binds to human red cell membranes suspended in isotonic NaCl at room temperature, with an apparent dissociation constant of 3×10−8m and maximum binding capacity of 1.8×107 molecules/cell. When about 1% of the melittin binding sites are occupied, cell lysis can be observed, and progressive, further increases in the fraction of the total sites occupied lead to progressively greater lysis in a graded manner. 50% lysis occurs when there are about 2×106 molecules bound to the cell membrane. For any particular extent of melittin binding, lysis proceeds rapidly during the first few minutes but then slows and stops so that no further lysis occurs after one hour of exposure of cells to melittin. The graded lysis of erythrocytes by melittin is due to complete lysis of some of the cells, since both the density and the hemoglobin content of surviving, intact cells in a suspension that has undergone graded melittin lysis are similar to the values observed in the same cells prior to the addition of melittin. The cells surviving graded melittin lysis have an increased Na and reduced K, proportional to the extent of occupation of the melittin binding sites. Like lysis, Na accumulation and K loss proceed rapidly during the first few minutes of exposure to melittin but then stops so that Na, K and hemoglobin content of the cells remain constant after the first hour. These kinetic characteristics of both lysis and cation movements suggest that melittin modifies the permeability of the red cell membrane only for the first few minutes after the start of the interaction. Direct observation of cells by Nomarsky optics revealed that they crenate, become swollen and lyse within 10 to 30 sec after these changes in morphology are first seen. Taken together, these results are consistent with the idea that melittin produces lysis of human red cells at room temperature by a colloid osmotic mechanism.

Key Words

melittin-induced osmotic lysis human red cells permeability increases 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bjerrum, P.J. 1979. Hemoglobin-depleted human erythrocyte ghosts: Characterization of morphology and transport functions.J. Membrane Biol. 48:43–67Google Scholar
  2. 2.
    Bowdler, A.J., Williams, R.H., Dougherty, R.M. 1984. Abrogation of calcium exclusion by erythrocytes under hypotonic stress.Scand. J. Haematol. 32:283–296Google Scholar
  3. 3.
    Brown, L.R., Lauterwein, J., Wüthrich, K. 1980. High resolution1H-NMR studies of self-aggregation of melittin in aqueous solution.Biochim. Biophys. Acta 622:231–244Google Scholar
  4. 4.
    Dannon, D., Marikovsky, Y. 1964. Determination of density distribution of red cell population.J. Lab. Clin. Med. 64:668–674Google Scholar
  5. 5.
    Davies, J.T., Rideal, E.K. 1963. Interfacial Phenomena. 2nd ed. Academic, New York and LondonGoogle Scholar
  6. 6.
    Dawson, C.R., Drake, A.F., Helliwell, J., Hider, R.C. 1978. The interaction of bee melittin with lipid bilayer membranes.Biochim. Biophys. Acta 510:75–86Google Scholar
  7. 7.
    DeGrado, W.F., Musso, G.F., Lieber, M., Kaiser, E.T., Kezdy, F.J. 1982. Kinetics and mechanism of hemolysis induced by melittin and by a synthetic melittin analogue.Biophys. J. 37:329–338Google Scholar
  8. 8.
    Dufourcq, J., Faucon, J.F. 1977. Intrinsic fluorescence study of lipid-protein interaction in membrane models. Binding of melittin, an amphipathic peptide, to phospholipid vesicles.Biochim. Biophys. Acta 467:1–11Google Scholar
  9. 9.
    Funder, J., Tosteson, D.C., Wieth, J.O. 1978. Effects of bicarbonate on lithium transport in human red cells.J. Gen. Physiol. 71:721–746Google Scholar
  10. 10.
    Habermann, E. 1972. Bee and wasp venoms.Science 177:314–322Google Scholar
  11. 11.
    Habermann, E., Ahnert-Hilger, G., Chatwal, G.S., Beress, L. 1981. Delayed hemolytic action of palytoxin. General charactertics.Biochim. Biophys. Acta 649:481–489Google Scholar
  12. 12.
    Hanke, W., C. Methfessel, C., Wilmesen, H.-U., Katz, E., Jung, G., Boheim, G. 1983. Mellitin and a chemically modified trichotoxin form alamethicin-type multi-state pores.Biochim. Biophys. Acta 727:108–114Google Scholar
  13. 13.
    Kemp, C., Klausner, R.D., Weinstein, J.N., Renswoude, J., van, Pincus, M., Blumenthal, R. 1982. Voltage-dependent trans-bilayer orientation of melittin.J. Biol. Chem. 257:2469–2476Google Scholar
  14. 14.
    Maulet, Y., Cox, J.A. 1983. Structural changes in melittin and calmodulin upon complex formation and their modulation by calcium.Biochemistry 22:5680–5686Google Scholar
  15. 15.
    Podo, F., Strom, R., Crifo, C., Zulauf, M. 1982. Dependence of melittin structure on its interaction with multivalent anions and with model membrane systems.Int. J. Peptide Protein Res. 19:514–527Google Scholar
  16. 16.
    Quay, S.C., Condie, C.C. 1983. Conformational studies of aqueous melittin: Thermodynamic parameters of the monomer-tetramer self association reaction.Biochemistry 22:695–700Google Scholar
  17. 17.
    Schoch, P., Sargent, D.F. 1980. Quantitative analysis of the binding of melittin to planar lipid bilayers allowing for the discrete-charge effect.Biochim. Biophys. Acta 602:234–247Google Scholar
  18. 18.
    Schröder, E., Lübke, K., Lehmann, M., Beetz, I. 1971. Hemolytic activity and action on the surface tension of aqueous solutions of synthetic melittins and their derivatives.Experientia 27:764–765Google Scholar
  19. 19.
    Sessa, G., Freer, J.H., Colacicco, G., Weismann, G. 1969. Interaction of a lytic peptide, melittin, with lipid membrane systems.J. Biol. Chem. 244:3575–3582Google Scholar
  20. 20.
    Sheetz, M.P., Singer, S.J. 1974. Biological membranes as bilayer couples. A molecular mechanism of drug-erythrocyte interactions.Proc. Natl. Acad. Sci. USA 71:4457–4461Google Scholar
  21. 21.
    Stankowski, S. 1983. Large-ligand adsorption to membranes. I. Linear ligands as a limiting case.Biochim. Biophys. Acta 735:341–351Google Scholar
  22. 22.
    Terwillinger, T.C., Weissman, L., Eisenberg, D. 1982. The structure of melittin in the form I crystals and its implication for melittin's lytic and surface activities.Biophys. J. 37:353–361Google Scholar
  23. 23.
    Tosteson, D.C. 1967. Electrolyte composition and transport in red blood cells.Fed. Proc. 26:1805–1812Google Scholar
  24. 24.
    Tosteson, M.T., Tosteson, D.C. 1981. The sting: Melittin forms channels in lipid bilayers.Biophys. J. 36:109–116Google Scholar
  25. 25.
    Tosteson, M.T., Tosteson, D.C. 1984. Activation and inactivation of melittin channels.Biophys. J. 45:112–114Google Scholar
  26. 26.
    Wilbrandt, W. 1941. Osmotische Natur sogenaunter nichtosmotischer Hämolysen (Kolloidosmotische Hämolyse).Pfluegers. Arch. Gesamte Physiol. 245:22Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • M. T. Tosteson
    • 1
  • S. J. Holmes
    • 1
  • M. Razin
    • 1
  • D. C. Tosteson
    • 1
  1. 1.Department of Physiology and BiophysicsHarvard Medical SchoolBoston

Personalised recommendations