Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 311, Issue 1, pp 95–103 | Cite as

Studies on rubescenslysin haemolysis

  • Ruth Seeger


Rubescenslysin from the edible mushroom Amanita rubescens (Pers. ex Fr.) Gray is an acidic protein that directly lyzes red cells. It is active even in the absence of Ca2+ and Mg2+ and its activity is not influenced by cysteine. — The concentration-response curve is steep. The time-course of haemolysis is characterized by a rather long lag phase; but after the onset of the haemolysis it proceeds quickly. The rate of haemolysis increases and the lag phase decreases with increasing lysin concentrations. The pH optimum is in the weekly acid range and the optimal temperature is 35°C. A pronounced increase in the neutral salt concentration inhibits the haemolysis; a self-inhibition is observed at higher lysin concentrations, particularly with preparations with a small degree of purity. — The haemolysis is of the osmotic type: a marked prelytic leakage of potassium as observed as well as a significant decrease of osmotic resistance on treatment of red cells with sublytic lysin concentrations. — Red cell sensitivity of various animal species decreases in the order: rat ≃ guinea pig = man = mouse ≃ dog ≃ rabbit > pig > cat > cattle = sheep; on the whole, species specifity is small. — The rubescenslysin haemolysis is inhibited by heavy metal salt (Cu2+>Fe2+>Zn2+>Cd2+>Mn2+>Ni2+) and by pretreatment of the erythrocytes with glutardialdehyde and wheat germ lectin > soya bean lectin. — Rubescenslysin is consumed on haemolysis and markedly inhibited by haemolysate (from bovine blood) as well as by red cell ghosts (from bovine and sheep blood). It is also inhibited by cholesterol and various phospholipids: sphingomyelin from bovine brain > sphingomyelin from chicken egg > phosphatidyl choline from egg yolk > phosphatidyl ethanolamine from dog brain; no inhibition is produced by phosphatidyl ethanolamine from bovine brain or soya bean. Of the synthetic phosphatidyl cholines dimyristoyl- has a stronger inhibitory effect than dipalmitoyl- and distearoyl-; on the other hand, dilauroyl- or dioleoyl- has no inhibitory effect. Human serum albumin or pretreatment of the erythrocytes with trypsin does not affect the haemolytic activity of rubescenslysin. — Surface activity of a rubescenslysin solution is greater than that of a serum albumin solution of the same concentration. — The results allow the conclusion that the cytolytic effect of rubescenslysin is due to a detergent activity of this protein.

Key words

Rubescenslysin Amanita rubescens Mushrooms Cytolysins Haemolysis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Avigad, L. S., Bernheimer, A. W.: Inhibition by zinc of hemolysis induced by bacterial and other cytolytic agents. Infect. Immun. 13, 1378–1381 (1976)Google Scholar
  2. Avigad, L. S., Bernheimer, A. W.: Inhibition of hemolysis by zinc and its reversal by Lxx-histidine. Infect. Immun. 19, 1101–1103 (1978)Google Scholar
  3. Bernheimer, A. W., Avigad, L. S.: A cholesterol-inhibitable cytolytic protein from the sea anemone Metridium senile. Biochim. Biophys. Acta 541, 96–106 (1978)Google Scholar
  4. Bernheimer, A. W., Avigad, L. S.: A cytolytic polypeptide from the edible mushroom Pleurotus ostreatus. Biochim. Biophys. Acta 585, 451–461 (1979)Google Scholar
  5. Capaldi, R. A. (ed.): Membrane proteins and their interactions with lipids. New York and Basel: Dekker 1977Google Scholar
  6. Crone, H. D.: Chemical modification of the haemolytic activity of the extracts from the box jellyfish Chironex fleckerii (cnidaria). Toxicon 14, 97–107 (1976)Google Scholar
  7. Dodge, J. D., Mitchell, C., Hanahan, D.: The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch. Biochem. Biophys. 100, 119–130 (1963)Google Scholar
  8. Faulstich, H., Weckauf, M.: Cytolysis of red cells mediated by phallolysin, a toxin binding to N-acetylglucosamine on the cell surface. Hoppe-Seylers Z. Physiol. Chem. 356, 1187–1189 (1975)Google Scholar
  9. Habermann, E.: Über die Wirkung tierischer Gifte auf Erythrozyten. Z. Ges. Exp. Med. 129, 436–464 (1958a)Google Scholar
  10. Habermann, E.: Zur Wirkung tierischer Gifte und von Lysocithin auf Grenzflächen. Z. Ges. Exp. Med. 130, 19–23 (1958b)Google Scholar
  11. Habermann, E.: Biochemie, Pharmakologie und Toxikologie der Inhaltsstoffe von Hymenopterengiften. Ergeb. Physiol. 60, 220–325 (1968)Google Scholar
  12. Jeljaszewicz, J., Szmigielski, S., Hryniewicz, W.: Biological effects of staphylococcal and streptococcal toxins. In: Bacterial toxins and cell membranes (Jeljaszewicz, J., Wadström, T., eds.), pp. 185–227. London, New York, San Francisco: Academic Press 1978Google Scholar
  13. Lehmann, V.: Haemolytic activity of Acinetobacter calcoaceticus. Acta Pathol. Microbiol. Scand. B. 79, 61–66 (1971)Google Scholar
  14. Lin, J.-Y., Jeng, T.-W., Chen, C.-C., Shi, G.-Y., Tung, T.-C.: Isolation of a new cardiotoxic protein from the edible mushroom Volvariella volvacea. Nature 246, 524–525 (1973)Google Scholar
  15. Lin, J.-Y., Lin, Y.-J., Chen, C.-C., Wu, H.-L., Shi, G.-Y., Jeng, T.-W.: Cardiotoxic protein from edible mushrooms. Nature 252, 235–237 (1974)Google Scholar
  16. Lin, J.-W., Wu, H.-L., Shi, G.-Y.: Toxicity of the cardiotoxic protein, flammutoxin, isolated from the edible mushroom Flammulina velutipes. Toxicon 13, 323–331 (1975)Google Scholar
  17. Lowry, O. W., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–273 (1951)Google Scholar
  18. Meyer, H. W., Winkelmann, H., Richter, W.: Digitonin induced alterations of the erythrocyte membrane as visible by freezefracturing. Exp. Pathol. (Jena) 16, 60–68 (1978)Google Scholar
  19. Montgomery, D. W., Don, L. K., Zukoski, C. F., Chvapil, M.: The effect of zinc and other metals on complement hemolysis of sheep red blood cells in vitro. Proc. Soc. Exp. Biol. Med. 145, 263–267 (1974)Google Scholar
  20. Odenthal, K. P., Mengs, U., Seeger, R.: Acute toxic effects of the haemolysin from Amanita rubescens after i.v. injection in rats. Naunyn-Schmiedeberg's Arch. Pharmacol. 294, R 24 (1976)Google Scholar
  21. Rouser, G., Nelson, G. J., Fleischer, S., Simon, G.: Lipid composition of animal cell membranes, organelles and organs. In: Biological membranes (Chapman, D., ed.), pp. 5–69. London and New York: Academic Press 1968Google Scholar
  22. Seeger, R.: Purification and some properties of the haemolysin from Amanita rubescens. Naunyn-Schmiedeberg's Arch. Pharmacol. 294, R 25 (1976)Google Scholar
  23. Seeger, R., Burkhardt, M.: The haemolytic effect of phallolysin. Naunyn-Schmiedeberg's Arch. Pharmacol. 293, 163–170 (1976)Google Scholar
  24. Seeger, R., Scharrer, H., Haupt, M.: Phallolysin, ein hochmolekulares Toxin aus Amanita phalloides. Experientia 29, 829 (1973)Google Scholar
  25. Takeda, Y., Ogiso, Y., Miwatani, T.: Effect of zinc ion on the hemolytic activity of thermostable direct hemolysin from Vibrio parahaemolyticus, streptolysin O, and Triton X-100. Infect. Immun. 17, 239–243 (1977)Google Scholar
  26. Tsukatani, H., Yamada, S., Fukuzawa, K., Hamaguchi, C.: Effect of lysolecithin on the systemic arterial blood pressure of anaesthetized rats. J. Pharm. Pharmacol. 31, 110–111 (1979)Google Scholar
  27. Weltzien, H. U.: Slow-reacting hemolytic phosphatides: Benzylated lysolecithins. Biochim. Biophys. Acta 311, 6–14 (1973)Google Scholar

Copyright information

© Springer-Verlag 1980

Authors and Affiliations

  • Ruth Seeger
    • 1
  1. 1.Institut für Pharmakologie und Toxikologie der Universität WürzburgWürzburgGermany

Personalised recommendations