This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hofmann, F.
Right arrow Articles by Aktories, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hofmann, F.
Right arrow Articles by Aktories, K.

 Previous Article  |  Next Article 

Infect Immun, March 1998, p. 1076-1081, Vol. 66, No. 3
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Chimeric Clostridial Cytotoxins: Identification of the N-Terminal Region Involved in Protein Substrate Recognition

Fred Hofmann, Christian Busch, and Klaus Aktories*

Institut für Pharmakologie und Toxikologie der Albert-Ludwigs-Universität Freiburg, D-79104 Freiburg, Germany

Received 18 September 1997/Returned for modification 27 October 1997/Accepted 24 November 1997

Clostridium sordellii lethal toxin is a member of the family of large clostridial cytotoxins that glucosylate small GTPases. In contrast to Clostridium difficile toxins A and B, which exclusively modify Rho subfamily proteins, C. sordellii lethal toxin also glucosylates Ras subfamily proteins. By deletion analysis and construction of chimeric fusion proteins of C. sordellii lethal toxin and C. difficile toxin B, we localized the enzyme activity of the lethal toxin to the N terminus of the holotoxin and identified the region involved in protein substrate specificity. The toxin fragment of the N-terminal 546 amino acid residues of C. sordellii lethal toxin glucosylated Rho and Ras subfamily proteins, as the holotoxin did. Deletion of a further 30 amino acid residues from the C terminus of this active fragment drastically reduced glucotransferase activity and blocked glucohydrolase activity. Exchange of amino acid residues 364 through 516 of lethal toxin for those in the active toxin B fragment (1 to 546) allowed glucosylation of Ras subfamily proteins. In contrast, the chimera with amino acids 1 to 364 from toxin B, 365 to 468 from lethal toxin, and 469 to 546 from toxin B exhibited markedly reduced modification of Ras subfamily proteins, whereas modification of Rac and Cdc42 was hardly changed. The data indicate that the region of amino acid residues 364 through 516 primarily defines the substrate specificity of C. sordellii lethal toxin.

* Corresponding author. Mailing address: Institut für Pharmakologie und Toxikologie der Albert-Ludwigs-Universität Freiburg, Hermann-Herder-Str. 5, 79104 Freiburg, Germany. Phone: 0761-2035301. Fax: 0761-2035311. E-mail: aktories{at}uni-freiburg.de.




This article has been cited by other articles:

  • Na, X., Kim, H., Moyer, M. P., Pothoulakis, C., LaMont, J. T. (2008). gp96 Is a Human Colonocyte Plasma Membrane Binding Protein for Clostridium difficile Toxin A. Infect. Immun. 76: 2862-2871 [Abstract] [Full Text]  
  • Jank, T., Pack, U., Giesemann, T., Schmidt, G., Aktories, K. (2006). Exchange of a Single Amino Acid Switches the Substrate Properties of RhoA and RhoD toward Glucosylating and Transglutaminating Toxins. J. Biol. Chem. 281: 19527-19535 [Abstract] [Full Text]  
  • Voth, D. E., Ballard, J. D. (2005). Clostridium difficile Toxins: Mechanism of Action and Role in Disease. Clin. Microbiol. Rev. 18: 247-263 [Abstract] [Full Text]  
  • Mesmin, B., Robbe, K., Geny, B., Luton, F., Brandolin, G., Popoff, M. R., Antonny, B. (2004). A Phosphatidylserine-binding Site in the Cytosolic Fragment of Clostridium sordellii Lethal Toxin Facilitates Glucosylation of Membrane-bound Rac and Is Required for Cytotoxicity. J. Biol. Chem. 279: 49876-49882 [Abstract] [Full Text]  
  • Voth, D. E., Qa'Dan, M., Hamm, E. E., Pelfrey, J. M., Ballard, J. D. (2004). Clostridium sordellii Lethal Toxin Is Maintained in a Multimeric Protein Complex. Infect. Immun. 72: 3366-3372 [Abstract] [Full Text]  
  • Spyres, L. M., Daniel, J., Hensley, A., Qa'Dan, M., Ortiz-Leduc, W., Ballard, J. D. (2003). Mutational Analysis of the Enzymatic Domain of Clostridium difficile Toxin B Reveals Novel Inhibitors of the Wild-Type Toxin. Infect. Immun. 71: 3294-3301 [Abstract] [Full Text]  
  • Humeau, Y., Popoff, M. R., Kojima, H., Doussau, F., Poulain, B. (2002). Rac GTPase Plays an Essential Role in Exocytosis by Controlling the Fusion Competence of Release Sites. J. Neurosci. 22: 7968-7981 [Abstract] [Full Text]  
  • Stevens, M. P., van Diemen, P. M., Frankel, G., Phillips, A. D., Wallis, T. S. (2002). Efa1 Influences Colonization of the Bovine Intestine by Shiga Toxin-Producing Escherichia coli Serotypes O5 and O111. Infect. Immun. 70: 5158-5166 [Abstract] [Full Text]  
  • Qa'Dan, M., Spyres, L. M., Ballard, J. D. (2001). pH-Enhanced Cytopathic Effects of Clostridium sordellii Lethal Toxin. Infect. Immun. 69: 5487-5493 [Abstract] [Full Text]  
  • Pothoulakis, C., Lamont, J. T. (2001). Microbes and Microbial Toxins: Paradigms for Microbial- Mucosal Interactions II. The integrated response of the intestine to Clostridium difficile toxins. Am. J. Physiol. Gastrointest. Liver Physiol. 280: G178-G183 [Abstract] [Full Text]  
  • Spyres, L. M., Qa'Dan, M., Meader, A., Tomasek, J. J., Howard, E. W., Ballard, J. D. (2001). Cytosolic Delivery and Characterization of the TcdB Glucosylating Domain by Using a Heterologous Protein Fusion. Infect. Immun. 69: 599-601 [Abstract] [Full Text]  
  • Busch, C., Schomig, K., Hofmann, F., Aktories, K. (2000). Characterization of the Catalytic Domain of Clostridium novyi Alpha-Toxin. Infect. Immun. 68: 6378-6383 [Abstract] [Full Text]  
  • Moncrief, J. S., Zheng, L., Neville, L. M., Lyerly, D. M. (2000). Genetic Characterization of Toxin A-Negative, Toxin B-Positive Clostridium difficile Isolates by PCR. J. Clin. Microbiol. 38: 3072-3075 [Abstract] [Full Text]  
  • Busch, C., Hofmann, F., Gerhard, R., Aktories, K. (2000). Involvement of a Conserved Tryptophan Residue in the UDP-Glucose Binding of Large Clostridial Cytotoxin Glycosyltransferases. J. Biol. Chem. 275: 13228-13234 [Abstract] [Full Text]  
  • Klapproth, J.-M. A., Scaletsky, I. C. A., McNamara, B. P., Lai, L.-C., Malstrom, C., James, S. P., Donnenberg, M. S. (2000). A Large Toxin from Pathogenic Escherichia coli Strains That Inhibits Lymphocyte Activation. Infect. Immun. 68: 2148-2155 [Abstract] [Full Text]  
  • Busch, C., Hofmann, F., Selzer, J., Munro, S., Jeckel, D., Aktories, K. (1998). A Common Motif of Eukaryotic Glycosyltransferases Is Essential for the Enzyme Activity of Large Clostridial Cytotoxins. J. Biol. Chem. 273: 19566-19572 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»