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. 1995 Apr;63(4):1188–1194. doi: 10.1128/iai.63.4.1188-1194.1995

Binding, oligomerization, and pore formation by streptolysin O in erythrocytes and fibroblast membranes: detection of nonlytic polymers.

I Walev 1, M Palmer 1, A Valeva 1, U Weller 1, S Bhakdi 1
PMCID: PMC173133  PMID: 7890371

Abstract

Streptolysin O (SLO) is a representative of the family of cholesterol-binding cytolysins that form large pores in target cell membranes. Aggregation of the toxin to polymeric structures is required for pore formation. However, it is not known whether, vice versa, polymers may under certain circumstances remain nonfunctional, and whether this might be the cause underlying the relative resistance of certain cells towards toxin action. In the present study, we applied radioiodinated, functionally active SLO to human, rabbit, and mouse erythrocytes and to human fibroblasts and keratinocytes. Binding and polymerization were quantified and correlated with membrane damage. At low toxin concentrations, human and rabbit but not mouse erythrocytes were lysed, but binding and polymerization of SLO were essentially identical in all cases. Nonlytic polymers were also detected on human fibroblasts and keratinocytes treated with subcytotoxic concentrations of SLO, and quantitative estimates indicated that nonpermeabilized cells could carry hundreds of polymers on their surface. When applied at low concentrations to fibroblasts, much of the toxin remained in monomer form and was subsequently shed from the cells. This was shown by monitoring the fate of radioiodinated toxin and also by using a sensitive cell enzyme-linked immunosorbent assay that permitted immunological detection of surface-exposed SLO. Thus, relative resistance of cells towards the permeabilizing action of SLO may be due to their ability to tolerate formation of a limited number of SLO polymers and to shedding of nonoligomerized toxin from their surface.

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Selected References

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  1. Alouf J. E. Streptococcal toxins (streptolysin O, streptolysin S, erythrogenic toxin). Pharmacol Ther. 1980;11(3):661–717. doi: 10.1016/0163-7258(80)90045-5. [DOI] [PubMed] [Google Scholar]
  2. Alving C. R., Habig W. H., Urban K. A., Hardegree M. C. Cholesterol-dependent tetanolysin damage to liposomes. Biochim Biophys Acta. 1979 Feb 20;551(1):224–228. doi: 10.1016/0005-2736(79)90368-7. [DOI] [PubMed] [Google Scholar]
  3. Bhakdi S., Grimminger F., Suttorp N., Walmrath D., Seeger W. Proteinaceous bacterial toxins and pathogenesis of sepsis syndrome and septic shock: the unknown connection. Med Microbiol Immunol. 1994 Jul;183(3):119–144. doi: 10.1007/BF00196048. [DOI] [PubMed] [Google Scholar]
  4. Bhakdi S., Roth M., Sziegoleit A., Tranum-Jensen J. Isolation and identification of two hemolytic forms of streptolysin-O. Infect Immun. 1984 Nov;46(2):394–400. doi: 10.1128/iai.46.2.394-400.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bhakdi S., Tranum-Jensen J. Damage to cell membranes by pore-forming bacterial cytolysins. Prog Allergy. 1988;40:1–43. [PubMed] [Google Scholar]
  6. Bhakdi S., Tranum-Jensen J., Sziegoleit A. Mechanism of membrane damage by streptolysin-O. Infect Immun. 1985 Jan;47(1):52–60. doi: 10.1128/iai.47.1.52-60.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bhakdi S., Weller U., Walev I., Martin E., Jonas D., Palmer M. A guide to the use of pore-forming toxins for controlled permeabilization of cell membranes. Med Microbiol Immunol. 1993 Sep;182(4):167–175. doi: 10.1007/BF00219946. [DOI] [PubMed] [Google Scholar]
  8. Buckingham L., Duncan J. L. Approximate dimensions of membrane lesions produced by streptolysin S and streptolysin O. Biochim Biophys Acta. 1983 Mar 23;729(1):115–122. doi: 10.1016/0005-2736(83)90462-5. [DOI] [PubMed] [Google Scholar]
  9. Cowell J. L., Bernheimer A. W. Role of cholesterol in the action of cereolysin on membranes. Arch Biochem Biophys. 1978 Oct;190(2):603–610. doi: 10.1016/0003-9861(78)90316-8. [DOI] [PubMed] [Google Scholar]
  10. Cowell J. L., Kim K. S., Bernheimer A. W. Alteration by cereolysin of the structure of cholesterol-containing membranes. Biochim Biophys Acta. 1978 Feb 21;507(2):230–241. doi: 10.1016/0005-2736(78)90419-4. [DOI] [PubMed] [Google Scholar]
  11. Domann E., Chakraborty T. Nucleotide sequence of the listeriolysin gene from a Listeria monocytogenes serotype 1/2a strain. Nucleic Acids Res. 1989 Aug 11;17(15):6406–6406. doi: 10.1093/nar/17.15.6406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dourmashkin R. R., Rosse W. F. Morphologic changes in the membranes of red blood cells undergoing hemolysis. Am J Med. 1966 Nov;41(5):699–710. doi: 10.1016/0002-9343(66)90031-3. [DOI] [PubMed] [Google Scholar]
  13. Duncan J. L., Schlegel R. Effect of streptolysin O on erythrocyte membranes, liposomes, and lipid dispersions. A protein-cholesterol interaction. J Cell Biol. 1975 Oct;67(1):160–174. doi: 10.1083/jcb.67.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Geoffroy C., Alouf J. E. Interaction of alveolysin A sulfhydryl-activated bacterial cytolytic toxin with thiol group reagents and cholesterol. Toxicon. 1982;20(1):239–241. doi: 10.1016/0041-0101(82)90208-2. [DOI] [PubMed] [Google Scholar]
  15. Geoffroy C., Mengaud J., Alouf J. E., Cossart P. Alveolysin, the thiol-activated toxin of Bacillus alvei, is homologous to listeriolysin O, perfringolysin O, pneumolysin, and streptolysin O and contains a single cysteine. J Bacteriol. 1990 Dec;172(12):7301–7305. doi: 10.1128/jb.172.12.7301-7305.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hugo F., Reichwein J., Arvand M., Krämer S., Bhakdi S. Use of a monoclonal antibody to determine the mode of transmembrane pore formation by streptolysin O. Infect Immun. 1986 Dec;54(3):641–645. doi: 10.1128/iai.54.3.641-645.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Iwamoto M., Nakamura M., Mitsui K., Ando S., Ohno-Iwashita Y. Membrane disorganization induced by perfringolysin O (theta-toxin) of Clostridium perfringens--effect of toxin binding and self-assembly on liposomes. Biochim Biophys Acta. 1993 Nov 21;1153(1):89–96. doi: 10.1016/0005-2736(93)90279-9. [DOI] [PubMed] [Google Scholar]
  18. Iwamoto M., Ohno-Iwashita Y., Ando S. Effect of isolated C-terminal fragment of theta-toxin (perfringolysin O) on toxin assembly and membrane lysis. Eur J Biochem. 1990 Nov 26;194(1):25–31. doi: 10.1111/j.1432-1033.1990.tb19422.x. [DOI] [PubMed] [Google Scholar]
  19. Johnson M. K., Geoffroy C., Alouf J. E. Binding of cholesterol by sulfhydryl-activated cytolysins. Infect Immun. 1980 Jan;27(1):97–101. doi: 10.1128/iai.27.1.97-101.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kehoe M. A., Miller L., Walker J. A., Boulnois G. J. Nucleotide sequence of the streptolysin O (SLO) gene: structural homologies between SLO and other membrane-damaging, thiol-activated toxins. Infect Immun. 1987 Dec;55(12):3228–3232. doi: 10.1128/iai.55.12.3228-3232.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kehoe M., Timmis K. N. Cloning and expression in Escherichia coli of the streptolysin O determinant from Streptococcus pyogenes: characterization of the cloned streptolysin O determinant and demonstration of the absence of substantial homology with determinants of other thiol-activated toxins. Infect Immun. 1984 Mar;43(3):804–810. doi: 10.1128/iai.43.3.804-810.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mengaud J., Chenevert J., Geoffroy C., Gaillard J. L., Cossart P. Identification of the structural gene encoding the SH-activated hemolysin of Listeria monocytogenes: listeriolysin O is homologous to streptolysin O and pneumolysin. Infect Immun. 1987 Dec;55(12):3225–3227. doi: 10.1128/iai.55.12.3225-3227.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mengaud J., Vicente M. F., Chenevert J., Pereira J. M., Geoffroy C., Gicquel-Sanzey B., Baquero F., Perez-Diaz J. C., Cossart P. Expression in Escherichia coli and sequence analysis of the listeriolysin O determinant of Listeria monocytogenes. Infect Immun. 1988 Apr;56(4):766–772. doi: 10.1128/iai.56.4.766-772.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mitsui K., Saeki Y., Hase J. Effects of cholesterol evulsion on susceptibility to perfringolysin O of human erythrocytes. Biochim Biophys Acta. 1982 Apr 7;686(2):177–181. doi: 10.1016/0005-2736(82)90110-9. [DOI] [PubMed] [Google Scholar]
  25. Mitsui K., Sekiya T., Okamura S., Nozawa Y., Hase J. Ring formation of perfringolysin O as revealed by negative stain electron microscopy. Biochim Biophys Acta. 1979 Dec 12;558(3):307–313. doi: 10.1016/0005-2736(79)90265-7. [DOI] [PubMed] [Google Scholar]
  26. Niedermeyer W. Interaction of streptolysin-O with biomembranes: kinetic and morphological studies on erythrocyte membranes. Toxicon. 1985;23(3):425–439. doi: 10.1016/0041-0101(85)90026-1. [DOI] [PubMed] [Google Scholar]
  27. Ohno-Iwashita Y., Iwamoto M., Mitsui K., Ando S., Nagai Y. Protease-nicked theta-toxin of Clostridium perfringens, a new membrane probe with no cytolytic effect, reveals two classes of cholesterol as toxin-binding sites on sheep erythrocytes. Eur J Biochem. 1988 Sep 1;176(1):95–101. doi: 10.1111/j.1432-1033.1988.tb14255.x. [DOI] [PubMed] [Google Scholar]
  28. Pinkney M., Beachey E., Kehoe M. The thiol-activated toxin streptolysin O does not require a thiol group for cytolytic activity. Infect Immun. 1989 Aug;57(8):2553–2558. doi: 10.1128/iai.57.8.2553-2558.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Prigent D., Alouf J. E. Interaction of steptolysin O with sterols. Biochim Biophys Acta. 1976 Aug 16;443(2):288–300. doi: 10.1016/0005-2736(76)90511-3. [DOI] [PubMed] [Google Scholar]
  30. Saunders F. K., Mitchell T. J., Walker J. A., Andrew P. W., Boulnois G. J. Pneumolysin, the thiol-activated toxin of Streptococcus pneumoniae, does not require a thiol group for in vitro activity. Infect Immun. 1989 Aug;57(8):2547–2552. doi: 10.1128/iai.57.8.2547-2552.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shany S., Bernheimer A. W., Grushoff P. S., Kim K. S. Evidence for membrane cholesterol as the common binding site for cereolysin, streptolysin O and saponin. Mol Cell Biochem. 1974 May 30;3(3):179–186. doi: 10.1007/BF01686643. [DOI] [PubMed] [Google Scholar]
  32. Thelestam M., Möllby R. Interaction of streptolysin O from Streptococcus pyogenes and theta-toxin from Clostridium perfringens with human fibroblasts. Infect Immun. 1980 Sep;29(3):863–872. doi: 10.1128/iai.29.3.863-872.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Thelestam M., Möllby R. Survival of cultured cells after functional and structural disorganization of plasma membrane by bacterial haemolysins and phospholipases. Toxicon. 1983;21(6):805–815. doi: 10.1016/0041-0101(83)90069-7. [DOI] [PubMed] [Google Scholar]
  34. Tweten R. K. Nucleotide sequence of the gene for perfringolysin O (theta-toxin) from Clostridium perfringens: significant homology with the genes for streptolysin O and pneumolysin. Infect Immun. 1988 Dec;56(12):3235–3240. doi: 10.1128/iai.56.12.3235-3240.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Walev I., Martin E., Jonas D., Mohamadzadeh M., Müller-Klieser W., Kunz L., Bhakdi S. Staphylococcal alpha-toxin kills human keratinocytes by permeabilizing the plasma membrane for monovalent ions. Infect Immun. 1993 Dec;61(12):4972–4979. doi: 10.1128/iai.61.12.4972-4979.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Walker J. A., Allen R. L., Falmagne P., Johnson M. K., Boulnois G. J. Molecular cloning, characterization, and complete nucleotide sequence of the gene for pneumolysin, the sulfhydryl-activated toxin of Streptococcus pneumoniae. Infect Immun. 1987 May;55(5):1184–1189. doi: 10.1128/iai.55.5.1184-1189.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wannamaker L. W. Streptococcal toxins. Rev Infect Dis. 1983 Sep-Oct;5 (Suppl 4):S723–S732. doi: 10.1093/clinids/5.supplement_4.s723. [DOI] [PubMed] [Google Scholar]
  38. Watson K. C., Kerr E. J. Sterol structural requirements for inhibition of streptolysin O activity. Biochem J. 1974 Apr;140(1):95–98. doi: 10.1042/bj1400095. [DOI] [PMC free article] [PubMed] [Google Scholar]

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