| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print July 9, 2001
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261
Corresponding Author: bamcc{at}pitt.edu
Since most in vitro studies exploring the action of Clostridium perfringens enterotoxin (CPE) utilize either Vero cells or CaCo-2 cells, the current study directly compared the CPE responsiveness of those two cell lines. When CPE-treated in suspension, both CaCo-2 and Vero cells formed SDS-resistant, CPE-containing complexes of ~135, ~155, and ~200kDa. However, when confluent Transwell“ cultures of either cell line were CPE-treated for 20 min only the ~155kDa complex formed. Since those Transwell“ cultures also exhibited significant 86Rb release, ~155kDa complex formation is sufficient to cause CPE-induced cytotoxicity. Several differences in CPE responsiveness between the two cell lines were also detected; i) CaCo-2 cells were more sensitive when CPE-treated on their basal surface, while Vero cells were more sensitive when CPE-treated on their apical surface; those sensitivity differences correlated with CPE binding differences between the apical vs. basolateral surfaces of these two cell lines, ii) CPE-treated Vero cells released 86Rb into both Transwell“ chambers, while CaCo-2 cells released 86Rb only into the CPE-containing Transwell“ chamber, and iii) Vero cells express the tight junction (TJ) protein occludin, but (unlike CaCo-2 cells) cannot form TJs. The ability of TJs to affect CPE responsiveness is supported by the similar effects of CPE on Transwell“ cultures of CaCo-2 cells and MDCK cells, another polarized cell forming TJs. Confluent CaCo-2 Transwell“ cultures CPE-treated for >1h formed the ~200kDa CPE complex (which also contains occludin), exhibited morphologic damage, and had occludin removed from their TJs. Collectively these results identify CPE as a bifunctional toxin that, in confluent polarized cells, first exerts a cytotoxic effect mediated by the ~155kDa complex. Resultant damage then provides CPE access to TJs, leading to ~200kDa complex formation, internalization of some TJ proteins, and TJ damage that may increase paracellular permeability and thereby contribute to the diarrheal symptoms of CPE-induced gastrointestinal disease.
J. Biol. Chem, 10.1074/jbc.M104200200
Submitted on May 9, 2001
Revised on June 21, 2001
Accepted on July 9, 2001
Comparative biochemical and immunocytochemical studies reveal differences in the effects of Clostridium perfringens enterotoxin on polarized CaCo-2 cells versus Vero cells
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  J. A. Caserta, M. L. Hale, M. R. Popoff, B. G. Stiles, and B. A. McClane Evidence that Membrane Rafts Are Not Required for the Action of Clostridium perfringens Enterotoxin Infect. Immun., December 1, 2008; 76(12): 5677 - 5685. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G. Smedley III, J. Saputo, J. C. Parker, M. E. Fernandez-Miyakawa, S. L. Robertson, B. A. McClane, and F. A. Uzal Noncytotoxic Clostridium perfringens Enterotoxin (CPE) Variants Localize CPE Intestinal Binding and Demonstrate a Relationship between CPE-Induced Cytotoxicity and Enterotoxicity Infect. Immun., August 1, 2008; 76(8): 3793 - 3800. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G. Smedley III, F. A. Uzal, and B. A. McClane Identification of a Prepore Large-Complex Stage in the Mechanism of Action of Clostridium perfringens Enterotoxin Infect. Immun., May 1, 2007; 75(5): 2381 - 2390. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Amimoto, T. Noro, E. Oishi, and M. Shimizu A novel toxin homologous to large clostridial cytotoxins found in culture supernatant of Clostridium perfringens type C Microbiology, April 1, 2007; 153(4): 1198 - 1206. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E. Fernandez Miyakawa, V. Pistone Creydt, F. A. Uzal, B. A. McClane, and C. Ibarra Clostridium perfringens Enterotoxin Damages the Human Intestine In Vitro Infect. Immun., December 1, 2005; 73(12): 8407 - 8410. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Masuyama, M. Kondoh, H. Seguchi, A. Takahashi, M. Harada, M. Fujii, H. Mizuguchi, Y. Horiguchi, and Y. Watanabe Role of N-Terminal Amino Acids in the Absorption-Enhancing Effects of the C-Terminal Fragment of Clostridium perfringens Enterotoxin J. Pharmacol. Exp. Ther., August 1, 2005; 314(2): 789 - 795. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G. Smedley III and B. A. McClane Fine Mapping of the N-Terminal Cytotoxicity Region of Clostridium perfringens Enterotoxin by Site-Directed Mutagenesis Infect. Immun., December 1, 2004; 72(12): 6914 - 6923. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Chakrabarti, X. Zhou, and B. A. McClane Death Pathways Activated in CaCo-2 Cells by Clostridium perfringens Enterotoxin Infect. Immun., August 1, 2003; 71(8): 4260 - 4270. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
