SpringerLink
Log in

Characterization of Vibrio cholerae El Tor cytolysin as an oligomerizing pore-forming toxin

  • Original Investigation
  • Published:
Medical Microbiology and Immunology Aims and scope Submit manuscript

Abstract

V. cholerae El Tor cytolysin is a secreted, water-soluble protein of M r 60,000 that may be relevant to the pathogenesis of acute diarrhea. In this communication, we demonstrate that the toxin binds to and oligomerizes in target membranes to form SDS-stable aggregates of M r 200000–250000 that generate small transmembrane pores. Pores formed in erythrocytes were approximately 0.7 nm in size, as demonstrated by osmotic protection experiments. Binding was shown to occur in a temperature-independent manner preceding the temperature-dependent oligomerization step. Pores were also shown to be formed in L929 and HEp-2 cells, human fibroblasts and keratinocytes, albeit with highly varying efficacy. At neutral pH and in the presence of serum, human fibroblasts were able to repair a limited number of lesions. The collective data identify V. cholerae El Tor cytolysin as an oligomerizing toxin that damages cells by creating small transmembrane pores.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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. Alm RA, Stroeher UH, Manning PA (1988) Extracellular proteins of Vibio cholerae: nucleotide sequence of structural gene (hlyA) for the hemolysin of the hemolytic El Tor strain 017 and characterization of the hlyA mutation in the nonhemolytic classical strain 569B. Mol Microbiol 2:481–488

    Google Scholar 

  2. Baumann P, Furniss AL, Lee JV (1984) Genus I. Vibrio. In: Krieg NR, Holt JG (eds) Bergey's manual of systematic bacteriology, vol 1. Williams & Wilkins, Baltimore, pp 518–538

    Google Scholar 

  3. Bhakdi S, Tranum-Jensen J (1988) Damage to cell membranes by pore-forming bacterial cytolysins. Prog Allergy 40:1–13

    Google Scholar 

  4. Bhakdi S, Mackman N, Nicaud J-M, Holland IB (1986) Escherichia coli hemolysin may damage target cell membranes by generating transmembrane pores. Infect Immun 52:63–69

    Google Scholar 

  5. Bhakdi S, Grimminger F, Suttorp N, Walmrath D, Seeger W (1994) Proteinaceous bacterial toxins and pathogenesis of sepsis syndrome and septic shock: the unknown connection. Med Microbiol Immunol 183:119–144

    Google Scholar 

  6. Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein, utilizing the principle of protein-dye binding. Anal Biochem 72:248–257

    Article  CAS  PubMed  Google Scholar 

  7. Garland WJ, Buckley JT (1988) The cytolytic toxin aerolysin must aggregate to disrupt erythrocyte, and aggregation is stimulated by human glycophorin. Infect Immun 56:1249–1253

    Google Scholar 

  8. Gelbart SM, Prabhudesari MM (1986) Vibrio cholerae non-01 cellulitis. Arch Pathol Lab Med 110:1182–1183

    Google Scholar 

  9. Geoffroy C, Gaillard J-L, Alouf JE, Berche P (1987) Purification, characterization and toxicity of the sulfhydryl-activated hemolysin listeriolysin O from Listeria monocytogenes. Infect Immun 55:1641–1646

    Google Scholar 

  10. Hall RH, Drasar BS (1990) Vibrio cholerae HlyA hemolysin is processed by proteolysis. Infect Immun 58:3375–3379

    Google Scholar 

  11. Honda T, Finkelstein RA (1979) Purification and characterization of a hemolysin produced by Vibrio cholerae biotype El Tor: another toxic substance produced by cholera vibrios. Infect Immun 26:1020–1027

    Google Scholar 

  12. Hughes JM, Hollis DG, Gangarosa EJ, Neaver RE (1978) Non-cholera Vibrio infections in the United States: clinical, epidemiologic and laboratory features. Ann Intern Med 88:602–606

    Google Scholar 

  13. Ichinose Y, Yamamoto K, Nakasone N, Tanabe MJ, Takeda T, Miwatani T, Iwanaga M (1987) Enterotoxicity of El Tor-like hemolysin of non-01 Vibrio cholerae. Infect Immun 55:1090–1093

    Google Scholar 

  14. Krasilnikov OV, Muratkhodjaev JN, Zitzer AO (1992) The mode of action of Vibrio cholerae cytolysin. The influences of both erythrocytes and planar lipid bilayers. Biochim Biophys Acta 1111:7–16

    Google Scholar 

  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    PubMed  Google Scholar 

  16. Lee MH, Leu HS, Huang SH (1993) Bacteremic cellulitis caused by non-01 Vibrio cholerae: report of a case. J Formos Med Assoc 92:472–474

    Google Scholar 

  17. Levine MM, Kaper JB, Herrington D, Losonsky G, Morris J, Clements ML, Black RE, Tall B, Hall R (1988) Volunteer studies of deletion mutants of Vibrio cholerae 01 prepared by recombinant techniques. Infect Immun 56:161–167

    Google Scholar 

  18. McCardell BA, Madden JM, Shah DB (1985) Isolation and characterization of a cytolysin produced by Vibrio cholerae sero- group non-01. Can J Microbiol 31:711–720

    Google Scholar 

  19. Michalsky J, Galen JE, Fasano A, Kaper JB (1993) CVD 110, an attenuated Vibrio cholerae 01 El Tor live oral vaccine strain. Infect Immun 61:4462–4468

    Google Scholar 

  20. Miyake M, Honda T, Miwatani T (1989) Effects of divalent cations and saccharides on Vibrio metschnikovii cytolysin-induced hemolysis of rabbit erythrocytes. Infect Immun 57:158–163

    Google Scholar 

  21. Morris JG Jr (1990) Non-01 group 1 Vibrio cholerae: a look at the epidemiology of an occasional pathogen. Epidemiol Rev 12:179–191

    Google Scholar 

  22. Naidu LS, Bakerman PR, Saubolle MA, Lewis K (1993) Vibrio cholerae non-01 meningitis in an infant. Pediatr Infect Dis J 12:879–881

    Google Scholar 

  23. Newman C, Shepherd M, Woodard MD, Chopra AK, Tyring SK (1993) Fatal septicemia and bullae caused by non-01 Vibrio cholerae. J Am Acad Dermatol 29:909–912

    Google Scholar 

  24. Nishibuchi M, Kaper JB (1985) Nucleotide sequence of the thermostable direct hemolysin of Vibrio parahaemolyticus. J Bacteriol 162:558

    Google Scholar 

  25. Rader AE, Murphy JR (1988) Nucleotide sequences and comparison of the hemolysin determinants of Vibrio cholerae El Tor RV79 (Hly+) and RV79 (Hly-) and classical 569 B (Hly-). Infect Immun 56:1414–1419

    Google Scholar 

  26. Safrin S, Morris JG, Adams M, Pons V, Jacobs R, Conte JE (1988) Non-01 Vibrio cholerae bacteremia: a case report and review. Rev Infect Dis 10:1012–1017

    Google Scholar 

  27. Scherrer R, Gerhardt P (1971) Molecular sieving by the Bacillus megaterium cell wall and protoplast. J Bacteriol 107: 718–735

    Google Scholar 

  28. Shelton CH, Martino RL, Ramsey KM (1993) Recurrent non-01 Vibrio cholerae bacteremia in a patient with multiple myeloma. Cancer 72:105–107

    Google Scholar 

  29. Shinoda S, Ichida K, Oh E-G, Sasahara K, Miyoshi S, Chowdhury M, Yasuda T (1993) Studies on hemolytic action of a hemolysin produced by Vibrio mumicus. Microbiol Immunol 327:405–409

    Google Scholar 

  30. Walev I, Martin E, Jonas D, Mohamadzadeh M, Müller-Klieser W, Kunz L, Bhakdi S (1993) Staphylococcal alpha-toxin kills human keratinoytes by permeabilizing the plasma membrane for monovalent ions. Infect Immun 61:4972–4979

    CAS  PubMed  Google Scholar 

  31. Walev I, Palmer M, Martin E, Jonas D, Weller U, Höhn-Bentz H, Husmann M, Bhakdi S (1994) Recover of human fibroblasts from attack by the pore-forming I-toxin of Staphylococcus aureus. Microb Pathog 17:187–201

    Article  CAS  PubMed  Google Scholar 

  32. Williams SG, Attridge SR, Manning PA (1993) The transcriptional activator HlyU of Vibrio cholerae: nucleotide sequence and role in virulence gene expression. Mol Microbiol 9:751–760

    Google Scholar 

  33. Yamamoto K, Al-Omani M, Honda T, Takeda J, Miwatani T (1984) Non-01 Vibrio cholerae hemolysin: purification, partial characterization and immunological relatedness to El Tor hemolysin. Infect Immun 45:192–196

    Google Scholar 

  34. Yamamoto K, Ichinose J, Nakasone N, Tanabe M, Nagahama M, Sakurai J, Iwanaga M (1986) Identity of hemolysins produced by Vibrio cholerae non-01 and V. cholerae 01 biotype El Tor. Infect Immun 51:927–931

    Google Scholar 

  35. Yamamoto K, Ichinose J, Shinagawa H, Makino K, Nakata A, Iwanaga M, Honda T, Miwatani T (1990) Two-step processing for activation of the cytolysin/hemolysin of Vibrio cholerae 01 biotype El Tor: nucleotide sequence of the structural gene (hylA) and characterization of the processed products. Infect Immun 58:4106–4116

    Google Scholar 

  36. Yamamoto K, Wright A, Kaper JB, Morris JG (1990) The cytolysin gene of Vibrio vulnificus: sequence and relationship to the Vibrio cholerae El Tor or hemolysin gene. Infect Immun 58:2706–2709

    Google Scholar 

  37. Yamanaka H, Satoh T, Katsu T, Shinoda S (1987) Mechanism of hemolysis by V. vulnificus hemolysin. J Gen Microbiol 133:2859–2864

    Google Scholar 

  38. Zitzer AO, Nakisbekov NO, Li AV, Semiotrochev VL, Kiseliov YL, Muratkhodjaev JN, Krasilnikov OV, Ezepchuk YV (1993) Enterocytolysin from Vibrio cholerae non-01 (some properties and pore-forming activity). Zentralbl Bakteriol 279:494–504

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Medical Microbiology and Hygiene, Hochhaus am Augustusplatz, D-55101, Mainz, Germany

    Alexander Zitzer, Iwan Walev, Michael Palmer & Sucharit Bhakdi

Authors
  1. Alexander Zitzer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Iwan Walev
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael Palmer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sucharit Bhakdi
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zitzer, A., Walev, I., Palmer, M. et al. Characterization of Vibrio cholerae El Tor cytolysin as an oligomerizing pore-forming toxin. Med Microbiol Immunol 184, 37–44 (1995). https://doi.org/10.1007/BF00216788

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00216788

Key words

Search

Navigation