![]()
|
|
||||||||
J. Biol. Chem., Vol. 275, Issue 25, 18704-18711, June 23, 2000
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the The actin-ADP-ribosylating binary
Clostridium botulinum C2 toxin consists of two individual
proteins, the binding/translocation component C2II and the enzyme
component C2I. To elicit its cytotoxic action, C2II binds to a receptor
on the cell surface and mediates cell entry of C2I via
receptor-mediated endocytosis. Here we report that binding of C2II to
the surface of target cells requires cleavage of C2II by trypsin.
Trypsin cleavage causes oligomerization of the activated C2II (C2IIa)
to give SDS-stable heptameric structures, which exhibit a
characteristic annular or horseshoe shape and form channels in lipid
bilayer membranes. Cytosolic delivery of the enzyme component C2I is
blocked by bafilomycin but not by brefeldin A or nocodazole, indicating
uptake from an endosomal compartment and requirement of endosomal
acidification for cell entry. In the presence of C2IIa and C2I, short
term acidification of the extracellular medium (pH 5.4) allows C2I to
enter the cytosol directly. Our data indicate that entry of C2 toxin
into cells involves (i) activation of C2II by trypsin-cleavage, (ii)
oligomerization of cleaved C2IIa to heptamers, (iii) binding of the
C2IIa oligomers to the carbohydrate receptor on the cell surface and
assembly with C2I, (iv) receptor-mediated endocytosis of both C2
components into endosomes, and finally (v) translocation and release of
C2I into the cytosol after acidification of the endosomal compartment.
Cellular Uptake of Clostridium botulinum C2 Toxin
Requires Oligomerization and Acidification*
,
,
,
,

Institut für Pharmakologie und
Toxikologie der Albert-Ludwigs-Universität Freiburg,
Hermann-Herder-Str. 5, D-79104 Freiburg, Germany,
¶ Max-Delbrück-Zentrum für Molekulare Medizin,
Robert-Rössle-Str. 10, D-13125 Berlin, Germany,
Department
of Biophysical Chemistry, University of Groningen, Nijenborgh 4, NL-9747 AG Groningen, The Netherlands, and ** Lehrstuhl für
Biotechnologie, Theodor-Boveri-Institut (Biozentrum) der
Universität Würzburg, Am Hubland,
D-97074 Würzburg, Germany
*
This study was supported by the Deutsche
Forschungsgemeinschaft (Sonderforschungsbereich 388) and by the Fonds
der Chemischen Industrie.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.

To whom correspondence should be addressed: Institut für
Pharmakologie und Toxikologie, Hermann-Herder-Str. 5, D-79104 Freiburg, Germany. Tel.: 49-761-2035301; Fax: 49-761-2035311; E-mail:
aktories@uni-freiburg.de.
This article has been cited by other articles:
![]() |
T. Neumeyer, B. Schiffler, E. Maier, A. E. Lang, K. Aktories, and R. Benz Clostridium botulinum C2 Toxin: IDENTIFICATION OF THE BINDING SITE FOR CHLOROQUINE AND RELATED COMPOUNDS AND INFLUENCE OF THE BINDING SITE ON PROPERTIES OF THE C2II CHANNEL J. Biol. Chem., February 15, 2008; 283(7): 3904 - 3914. [Abstract] [Full Text] [PDF] |
||||
![]() |
S. Pust, H. Hochmann, E. Kaiser, G. von Figura, K. Heine, K. Aktories, and H. Barth A Cell-permeable Fusion Toxin as a Tool to Study the Consequences of Actin-ADP-ribosylation Caused by the Salmonella enterica Virulence Factor SpvB in Intact Cells J. Biol. Chem., April 6, 2007; 282(14): 10272 - 10282. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Giesemann, T. Jank, R. Gerhard, E. Maier, I. Just, R. Benz, and K. Aktories Cholesterol-dependent Pore Formation of Clostridium difficile Toxin A J. Biol. Chem., April 21, 2006; 281(16): 10808 - 10815. [Abstract] [Full Text] [PDF] |
||||
![]() |
C. Genisset, C. L. Galeotti, P. Lupetti, D. Mercati, D. A. G. Skibinski, S. Barone, R. Battistutta, M. de Bernard, and J. L. Telford A Helicobacter pylori Vacuolating Toxin Mutant That Fails To Oligomerize Has a Dominant Negative Phenotype Infect. Immun., March 1, 2006; 74(3): 1786 - 1794. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. Barth, K. Aktories, M. R. Popoff, and B. G. Stiles Binary Bacterial Toxins: Biochemistry, Biology, and Applications of Common Clostridium and Bacillus Proteins Microbiol. Mol. Biol. Rev., September 1, 2004; 68(3): 373 - 402. [Abstract] [Full Text] [PDF] |
||||
![]() |
G. Haug, K. Aktories, and H. Barth The Host Cell Chaperone Hsp90 Is Necessary for Cytotoxic Action of the Binary Iota-Like Toxins Infect. Immun., May 1, 2004; 72(5): 3066 - 3068. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. Vendeville, F. Rayne, A. Bonhoure, N. Bettache, P. Montcourrier, and B. Beaumelle HIV-1 Tat Enters T Cells Using Coated Pits before Translocating from Acidified Endosomes and Eliciting Biological Responses Mol. Biol. Cell, May 1, 2004; 15(5): 2347 - 2360. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. Fischer, K. Kohler, M. Fotin-Mleczek, and R. Brock A Stepwise Dissection of the Intracellular Fate of Cationic Cell-penetrating Peptides J. Biol. Chem., March 26, 2004; 279(13): 12625 - 12635. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Matsuzawa, A. Fukui, T. Kashimoto, K. Nagao, K. Oka, M. Miyake, and Y. Horiguchi Bordetella Dermonecrotic Toxin Undergoes Proteolytic Processing to Be Translocated from a Dynamin-related Endosome into the Cytoplasm in an Acidification-independent Manner J. Biol. Chem., January 23, 2004; 279(4): 2866 - 2872. [Abstract] [Full Text] [PDF] |
||||
![]() |
D. Blocker, K. Pohlmann, G. Haug, C. Bachmeyer, R. Benz, K. Aktories, and H. Barth Clostridium botulinum C2 Toxin: LOW pH-INDUCED PORE FORMATION IS REQUIRED FOR TRANSLOCATION OF THE ENZYME COMPONENT C2I INTO THE CYTOSOL OF HOST CELLS J. Biol. Chem., September 26, 2003; 278(39): 37360 - 37367. [Abstract] [Full Text] [PDF] |
||||
![]() |
G. Haug, J. Leemhuis, D. Tiemann, D. K. Meyer, K. Aktories, and H. Barth The Host Cell Chaperone Hsp90 Is Essential for Translocation of the Binary Clostridium botulinum C2 Toxin into the Cytosol J. Biol. Chem., August 22, 2003; 278(34): 32266 - 32274. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. Galle, A. Mameghani, S.-S. Bolz, S. Gambaryan, M. Gorg, T. Quaschning, U. Raff, H. Barth, S. Seibold, C. Wanner, et al. Oxidized LDL and its Compound Lysophosphatidylcholine Potentiate AngII-Induced Vasoconstriction by Stimulation of RhoA J. Am. Soc. Nephrol., June 1, 2003; 14(6): 1471 - 1479. [Abstract] [Full Text] [PDF] |
||||
![]() |
J.-C. Marvaud, B. G. Stiles, A. Chenal, D. Gillet, M. Gibert, L. A. Smith, and M. R. Popoff Clostridium perfringens Iota Toxin. MAPPING OF THE Ia DOMAIN INVOLVED IN DOCKING WITH Ib AND CELLULAR INTERNALIZATION J. Biol. Chem., November 8, 2002; 277(46): 43659 - 43666. [Abstract] [Full Text] [PDF] |
||||
![]() |
O. Knapp, R. Benz, M. Gibert, J. C. Marvaud, and M. R. Popoff Interaction of Clostridium perfringens Iota-Toxin with Lipid Bilayer Membranes. DEMONSTRATION OF CHANNEL FORMATION BY THE ACTIVATED BINDING COMPONENT Ib AND CHANNEL BLOCK BY THE ENZYME COMPONENT Ia J. Biol. Chem., February 15, 2002; 277(8): 6143 - 6152. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. Barth, R. Roebling, M. Fritz, and K. Aktories The Binary Clostridium botulinum C2 Toxin as a Protein Delivery System. IDENTIFICATION OF THE MINIMAL PROTEIN REGION NECESSARY FOR INTERACTION OF TOXIN COMPONENTS J. Biol. Chem., February 8, 2002; 277(7): 5074 - 5081. [Abstract] [Full Text] [PDF] |
||||
![]() |
D. Blocker, J. Behlke, K. Aktories, and H. Barth Cellular Uptake of the Clostridium perfringens Binary Iota-Toxin Infect. Immun., May 1, 2001; 69(5): 2980 - 2987. [Abstract] [Full Text] [PDF] |
||||
![]() |
J.-C. Marvaud, T. Smith, M. L. Hale, M. R. Popoff, L. A. Smith, and B. G. Stiles Clostridium perfringens Iota-Toxin: Mapping of Receptor Binding and Ia Docking Domains on Ib Infect. Immun., April 1, 2001; 69(4): 2435 - 2441. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. Barth, G. Pfeifer, F. Hofmann, E. Maier, R. Benz, and K. Aktories Low pH-induced Formation of Ion Channels by Clostridium difficile Toxin B in Target Cells J. Biol. Chem., March 30, 2001; 276(14): 10670 - 10676. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |