Hostname: page-component-7479d7b7d-t6hkb Total loading time: 0 Render date: 2024-07-11T12:00:02.528Z Has data issue: false hasContentIssue false
  • English
  • Français

Spontaneous release of cysteine proteinases but not of pore-forming peptides by viable Entamoeba histolytica

Published online by Cambridge University Press:  06 April 2009

M. Leippe ,
H. J. Sievertsen ,
E. Tannich  and
R. D. Horstmann
M. Leippe
Affiliation:
Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Germany
H. J. Sievertsen
Affiliation:
Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Germany
E. Tannich
Affiliation:
Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Germany
R. D. Horstmann
Affiliation:
Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Germany
Get access Rights & Permissions [Opens in a new window]

Summary

Invasive properties of pathogenic Entamoeba histolytica have been postulated to depend on the secretion or release of cysteine proteinases and pore-forming peptides (amoebapores) by trophozoites. To establish whether such toxic molecules are released by viable trophozoites or upon cellular disintegration, amoebae were maintained in various culture media, and activities in supernatants were monitored over time in correlation to cellular integrity. By measuring the release of the cytoplasmic marker enzyme NADP+-alcohol dehydrogenase, it became apparent that release of amoebapore was accompanied by cellular disintegration. In contrast, considerable quantities of cysteine proteinases were found to be present in culture supernatants also when amoebae remained intact. Treatment of amoebae with concanavalin A, bacterial lipo-polysaccharides or the calcium ionophore A23187 did not result in amoebapore secretion suggesting that here target cell contact is required as an essential stimulus.

Keywords

Entamoeba histolyticaamoebiasisamoebaporescysteine proteinasesmarker enzymeviability
Type
Research Article
Information
Parasitology , Volume 111 , Issue 5 , December 1995 , pp. 569 - 574
Copyright
Copyright © Cambridge University Press 1995

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

AndrÄ, J. & Leippe, M. (1994). Pore-forming peptide of Entamoeba histolytica: Significance of positively charged amino acid residues for its mode of action. FEBS Letters 354, 97–102.CrossRefGoogle ScholarPubMed
Arias-Negrete, S., Villagomez-Castro, J. C., Anaya-Velazquez, F. & Lira-Ortiz, R. (1991). Entamoeba histolytica: A simplified method to quantify its cytotoxicity. International Journal for Parasitology 21, 373–5.CrossRefGoogle ScholarPubMed
Bos, H. J. (1979). Entamoeba histolytica: Cytopathogenicity of intact amoebae and cell-free extracts; isolation and characterization of an intracellular toxin. Experimental Parasitology 47, 369–77.CrossRefGoogle ScholarPubMed
Cano-Mancera, R. & Lopez-Revilla, R. (1988). Maintenance of integrity, viability, and adhesion of Entamoeba histolytica trophozoites in different incubation media. Journal of Protozoology 35, 470–5.CrossRefGoogle ScholarPubMed
Diamond, L. S., Harlow, D. R. & Cunnick, C. C. (1978). A new medium for axenic cultivation of Entamoeba histolytica and other Entamoeba. Transactions of the Royal Society of Tropical Medicine and Hygiene 72, 431–2.CrossRefGoogle ScholarPubMed
Feingold, C., Bracha, R., Wexler, A. & Mirelman, D. (1985). Isolation, purification, and partial characterization of an enterotoxin from extracts of Entamoeba histolytica trophozoites. Infection and Immunity 48, 211–18.CrossRefGoogle ScholarPubMed
Gadasi, H. & Kobiler, D. (1983). Entamoeba histolytica: Correlation between virulence and content of proteolytic enzymes. Experimental Parasitology 55, 105–10.CrossRefGoogle ScholarPubMed
Ganz, T. (1987). Extracellular release of antimicrobial defensins by human polymorphonuclear leukocytes. Infection and Immunity 55, 568–71.CrossRefGoogle ScholarPubMed
Horstmann, R. D., Leippe, M. & Tannich, E. (1992). Recent progress in the molecular biology of Entamoeba histolytica. Tropical Medicine and Parasitology 43, 213–18.Google ScholarPubMed
Keene, W. E., Petitt, M. G., Allen, S. & McKerrow, J. H. (1986). The major neutral proteinase of Entamoeba histolytica. Journal of Experimental Medicine 163, 536–49.CrossRefGoogle ScholarPubMed
Keene, W. E., Hidalgo, M. E., Orozco, E. & McKerrow, J. H. (1990). Entamoeba histolytica: Correlation of the cytopathic effect of virulent trophozoites with secretion of a cysteine proteinase. Experimental Parasitology 71, 199–206.CrossRefGoogle ScholarPubMed
Kumar, A., Shen, P.-S., Descoteaux, S., Pohl, J., Bayley, G. & Samuelson, J. (1992). Cloning and expression of an NADP+-dependent alcohol dehydrogenase gene of Entamoeba histolytica. Proceedings of the National Academy of Sciences, USA 89, 10188–92.CrossRefGoogle ScholarPubMed
Leippe, M., Ebel, S., Schoenberger, O. L., Horstmann, R. D. & Müller-Eberhard, H. J. (1991). Pore-forming peptide of pathogenic Entamoeba histolytica. Proceedings of the National Academy of Sciences, USA 88, 7659–63.CrossRefGoogle ScholarPubMed
Leippe, M. & Müller-Eberhard, H. J. (1994). The pore-forming peptide of Entamoeba histolytica, the protozoan parasite causing human amoebiasis. Toxicology 87, 5–18.CrossRefGoogle ScholarPubMed
Leippe, M., AndrÄ, J. & Müller-Eberhard, H. J. (1994 a). Cytolytic and antibacterial activity of synthetic peptides derived from amoebapore, the pore-forming peptide of Entamoeba histolytica. Proceedings of the National Academy of Sciences, USA 91, 2602–6.CrossRefGoogle ScholarPubMed
Leippe, M., AndrÄ, J., Nickel, R., Tannich, E. & Müller-Eberhard, H. J. (1994 b). Amoebapores, a family of membranolytic peptides from the cytoplasmic granules of Entamoeba histolytica: isolation, primary structure, and pore formation in bacterial cytoplasmic membranes. Molecular Microbiology 14, 895–904.CrossRefGoogle ScholarPubMed
Lo, H.-S. & Chang, C.-J. (1982) Purification and properties of NADP-linked, alcohol dehydrogenase from Entamoeba histolytica. Journal of Parasitology 68, 372–7.CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 165–75.CrossRefGoogle ScholarPubMed
Luaces, A. L. & Barrett, A. J. (1988). Affinity purification and biochemical characterization of histolysin, the major cysteine proteinase of Entamoeba histolytica. The Biochemical Journal 250, 903–9.CrossRefGoogle ScholarPubMed
Lushbaugh, W. B., Hofbauer, A. F. & Pitmann, F. E. (1985). Entamoeba histolytica: purification of Cathepsin B. Experimental Parasitology 59, 328–36.CrossRefGoogle ScholarPubMed
Müller, F.-W., Franz, A. & Werries, E. (1988). Secretory hydrolases of Entamoeba histolytica. Journal of Protozoology 35, 291–5.CrossRefGoogle ScholarPubMed
Petri, W. A. & Ravdin, J. I. (1988). In vitro models of amebic pathogenesis. In Amebiasis – Human Infection by Entamoeba histolytica (ed. Ravdin, J. I.) pp. 191–204, New York: John Wiley & Sons.Google Scholar
Ravdin, J. I. (1989). Immunobiology of human infection by Entamoeba histolytica. Pathology and Immunopathology Research 8, 179–205.CrossRefGoogle ScholarPubMed
Reed, S. L., Keene, W. E. & McKerrow, J. H. (1989). Thiol proteinase expression and pathogenicity of Entamoeba histolytica. Journal of Clinical Microbiology 27, 2772–7.CrossRefGoogle ScholarPubMed
Scholze, H., Löhden-Bendinger, U., Müller, G. & Bakker-Grunwald, T. (1992). Subcellular distribution of amebapain, the major cysteine proteinase of Entamoeba histolytica. Archives of Medical Research 23, 105–8.Google ScholarPubMed
Scholze, H. & Tannich, E. (1994). Cysteine endopeptidases of Entamoeba histolytica. Methods in Enzymology 244, 512–22.CrossRefGoogle ScholarPubMed
Young, J. D.-E., Young, T. M., Lu, L. P., Unkeless, J. C. & Cohn, Z. A. (1982). Characterization of a membrane pore-forming protein from Entamoeba histolytica. Journal of Experimental Medicine 156, 1677–90.CrossRefGoogle ScholarPubMed