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Proteolysis of enteric cell villin by Entamoeba histolytica cysteine proteinases

(2003) JOURNAL OF BIOLOGICAL CHEMISTRY. 278(25). p.22650-22656
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Abstract
Invasive microorganisms efface enteric microvilli to establish intimate contact with the apical surface of enterocytes. To understand the molecular basis of this effacement in amebic colitis, we seeded Entamoeba histolytica trophozoites on top of differentiated human Caco-2 cell layers. Western blots of detergent lysates from such cocultures showed proteolysis of the actin-bundling protein villin within 1 min of direct contact of living trophozoites with enterocytes. Mixtures of separately prepared lysates excluded detergent colysis as the cause of villin proteolysis. Caspases were not responsible as evidenced by the lack of degradation of specific substrates and the failure of a specific caspase inhibitor to prevent villin proteolysis. A crucial role for amebic cysteine proteinases was shown by prevention of villin proteolysis and associated microvillar alterations through the treatment of trophozoites before coculture with synthetic inhibitors that completely blocked amebic cysteine proteinase activity on zymograms. Moreover, trophozoites of amebic strains pSA8 and SAW760 with strongly reduced cysteine proteinase activity showed a reduced proteolysis of villin in coculture with enteric cells. Salmonella typhimurium and enteropathogenic Escherichia coli disturb microvilli without villin proteolysis, indicating that the latter is not a consequence of the disturbance of microvilli. In conclusion, villin proteolysis is an early event in the molecular cross-talk between enterocytes and amebic trophozoites, causing a disturbance of microvilli.
Keywords
EXPRESSION, MECHANISMS, TROPHOZOITES, CATHEPSIN-B, CANCER-CELLS, ACTIN CYTOSKELETON, HOST-CELLS, IN-VITRO, ENTEROPATHOGENIC ESCHERICHIA-COLI, LIVER-ABSCESS FORMATION

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Chicago
Lauwaet, Tineke, Maria José Oliveira, Bert Callewaert, Georges De Bruyne, Xavier Saelens, Serge Ankri, Peter Vandenabeele, David Mirelman, Marcus Mareel, and Ancy Leroy. 2003. “Proteolysis of Enteric Cell Villin by Entamoeba Histolytica Cysteine Proteinases.” Journal of Biological Chemistry 278 (25): 22650–22656.
APA
Lauwaet, T., Oliveira, M. J., Callewaert, B., De Bruyne, G., Saelens, X., Ankri, S., Vandenabeele, P., et al. (2003). Proteolysis of enteric cell villin by Entamoeba histolytica cysteine proteinases. JOURNAL OF BIOLOGICAL CHEMISTRY, 278(25), 22650–22656.
Vancouver
1.
Lauwaet T, Oliveira MJ, Callewaert B, De Bruyne G, Saelens X, Ankri S, et al. Proteolysis of enteric cell villin by Entamoeba histolytica cysteine proteinases. JOURNAL OF BIOLOGICAL CHEMISTRY. 2003;278(25):22650–6.
MLA
Lauwaet, Tineke, Maria José Oliveira, Bert Callewaert, et al. “Proteolysis of Enteric Cell Villin by Entamoeba Histolytica Cysteine Proteinases.” JOURNAL OF BIOLOGICAL CHEMISTRY 278.25 (2003): 22650–22656. Print.
@article{211055,
  abstract     = {Invasive microorganisms efface enteric microvilli to establish intimate contact with the apical surface of enterocytes. To understand the molecular basis of this effacement in amebic colitis, we seeded Entamoeba histolytica trophozoites on top of differentiated human Caco-2 cell layers. Western blots of detergent lysates from such cocultures showed proteolysis of the actin-bundling protein villin within 1 min of direct contact of living trophozoites with enterocytes. Mixtures of separately prepared lysates excluded detergent colysis as the cause of villin proteolysis. Caspases were not responsible as evidenced by the lack of degradation of specific substrates and the failure of a specific caspase inhibitor to prevent villin proteolysis. A crucial role for amebic cysteine proteinases was shown by prevention of villin proteolysis and associated microvillar alterations through the treatment of trophozoites before coculture with synthetic inhibitors that completely blocked amebic cysteine proteinase activity on zymograms. Moreover, trophozoites of amebic strains pSA8 and SAW760 with strongly reduced cysteine proteinase activity showed a reduced proteolysis of villin in coculture with enteric cells. Salmonella typhimurium and enteropathogenic Escherichia coli disturb microvilli without villin proteolysis, indicating that the latter is not a consequence of the disturbance of microvilli. In conclusion, villin proteolysis is an early event in the molecular cross-talk between enterocytes and amebic trophozoites, causing a disturbance of microvilli.},
  author       = {Lauwaet, Tineke and Oliveira, Maria Jos{\'e} and Callewaert, Bert and De Bruyne, Georges and Saelens, Xavier and Ankri, Serge and Vandenabeele, Peter and Mirelman, David and Mareel, Marcus and Leroy, Ancy},
  issn         = {0021-9258},
  journal      = {JOURNAL OF BIOLOGICAL CHEMISTRY},
  keyword      = {EXPRESSION,MECHANISMS,TROPHOZOITES,CATHEPSIN-B,CANCER-CELLS,ACTIN CYTOSKELETON,HOST-CELLS,IN-VITRO,ENTEROPATHOGENIC ESCHERICHIA-COLI,LIVER-ABSCESS FORMATION},
  language     = {eng},
  number       = {25},
  pages        = {22650--22656},
  title        = {Proteolysis of enteric cell villin by Entamoeba histolytica cysteine proteinases},
  url          = {http://dx.doi.org/10.1074/jbc.M300142200},
  volume       = {278},
  year         = {2003},
}

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