HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 281, Issue 16, 10808-10815, April 21, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/16/10808    most recent M512720200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Giesemann, T. Articles by Aktories, K. Search for Related Content PubMed PubMed Citation Articles by Giesemann, T. Articles by Aktories, K. Social Bookmarking                      What's this?

Cholesterol-dependent Pore Formation of Clostridium difficile Toxin A^*

Torsten Giesemann, Thomas Jank, Ralf Gerhard, Elke Maier^¶, Ingo Just, Roland Benz^¶, and Klaus Aktories¹

From the Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Albert-Ludwigs-Universität Freiburg, D-79104 Freiburg, Germany, Institut für Toxikologie der Medizinischen Hochschule Hannover, D-30625 Hannover, Germany, and ^¶Lehrstuhl für Biotechnologie, Biozentrum, Am Hubland, D-97074 Würzburg, Germany

The large clostridial cytotoxins toxin A and toxin B from Clostridium difficile are major virulence factors known to cause antibiotic-associated diarrhea and pseudomembranous colitis. Both toxins mono-glucosylate and thereby inactivate small GTPases of the Rho family. Recently, it was reported that toxin B, but not toxin A, induces pore formation in membranes of target cells under acidic conditions. Here, we reassessed data on pore formation of toxin A in cells derived from human colon carcinoma. Treatment of ⁸⁶Rb⁺-loaded cells with native or recombinant toxin A resulted in an increased efflux of radioactive cations induced by an acidic pulse. The efficacy of pore formation was dependent on membrane cholesterol, since cholesterol depletion of membranes with methyl--cyclodextrin inhibited ⁸⁶Rb⁺ efflux, and cholesterol repletion reconstituted pore-forming activity of toxin A. Similar results were obtained with toxin B. Consistently, methyl--cyclodextrin treatment delayed intoxication of cells in a concentration-dependent manner. In black lipid membranes, toxin A induced ion-permeable pores only in cholesterol containing bilayers and at low pH. In contrast, release of glycosylphosphatidylinositol-anchored structures by phosphatidylinositol specific phospholipase C treatment did not reduce cell sensitivity toward toxins A and B. These data indicate that in colonic cells toxin A induces pore formation in an acidic environment (e.g. endosomes) similar to that reported for toxin B and suggest that pore formation by clostridial glucosylating toxins depends on the presence of cholesterol.

Received for publication, November 29, 2005 , and in revised form, February 13, 2006.

^* The study was supported by Deutsche Forschungsgemeinschaft Project Ak 6/16-1, Bundesministerium für Bildung und Forschung research group Klinische Infektiologie Freiburg, TP 1a, Deutsche Forschungsgemeinschaft Grant SFB 621 Project B5 (to R. G.), SFB 487 Project A5 (to R. B.), European Concerted Action Grant QLK-CT2001-01267 (to I. J.), and the Fonds der Chemischen Industrie. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Institut für Experimentelle und Klinische Pharmakologie und Toxikologie der Albert-Ludwigs-Universität Freiburg, Albert-Strasse 25, D-79104 Freiburg, Germany. Tel.: 49-761-2035301; Fax: 49-761-2035311; E-mail: Klaus.Aktories{at}pharmakol.uni-freiburg.de.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				X. He, X. Sun, J. Wang, X. Wang, Q. Zhang, S. Tzipori, and H. Feng Antibody-Enhanced, Fc Gamma Receptor-Mediated Endocytosis of Clostridium difficile Toxin A Infect. Immun., June 1, 2009; 77(6): 2294 - 2303. [Abstract] [Full Text] [PDF]

				M. Egerer, T. Giesemann, C. Herrmann, and K. Aktories Autocatalytic Processing of Clostridium difficile Toxin B: BINDING OF INOSITOL HEXAKISPHOSPHATE J. Biol. Chem., February 6, 2009; 284(6): 3389 - 3395. [Abstract] [Full Text] [PDF]

				T. Giesemann, M. Egerer, T. Jank, and K. Aktories Processing of Clostridium difficile toxins J. Med. Microbiol., June 1, 2008; 57(6): 690 - 696. [Abstract] [Full Text] [PDF]

				C. E. Soltani, E. M. Hotze, A. E. Johnson, and R. K. Tweten Structural elements of the cholesterol-dependent cytolysins that are responsible for their cholesterol-sensitive membrane interactions PNAS, December 18, 2007; 104(51): 20226 - 20231. [Abstract] [Full Text] [PDF]

				D. N. Atapattu and C. J. Czuprynski Mannheimia haemolytica Leukotoxin Binds to Lipid Rafts in Bovine Lymphoblastoid Cells and Is Internalized in a Dynamin-2- and Clathrin-Dependent Manner Infect. Immun., October 1, 2007; 75(10): 4719 - 4727. [Abstract] [Full Text] [PDF]

				T. Jank, T. Giesemann, and K. Aktories Rho-glucosylating Clostridium difficile toxins A and B: new insights into structure and function Glycobiology, April 1, 2007; 17(4): 15R - 22R. [Abstract] [Full Text] [PDF]