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 Nagahama, M.
Right arrow Articles by Sakurai, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nagahama, M.
Right arrow Articles by Sakurai, J.

 Previous Article  |  Next Article 

Journal of Bacteriology, April 2000, p. 2096-2103, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of the Enzymatic Component of Clostridium perfringens Iota-Toxin

Masahiro Nagahama, Yoshihiko Sakaguchi, Keiko Kobayashi, Sadayuki Ochi, and Jun Sakurai*

Department of Microbiology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Tokushima 770-8514, Japan

Received 4 October 1999/Accepted 20 January 2000

The iotaa component (ia) of Clostridium perfringens ADP ribosylates nonmuscle beta /gamma actin and skeletal muscle alpha -actin. Replacement of Arg-295 in ia with alanine led to a complete loss of NAD+-glycohydrolase (NADase) and ADP-ribosyltransferase (ARTase); that of the residue with lysine caused a drastic reduction in NADase and ARTase activities (<0.1% of the wild-type activities) but did not completely diminish them. Substitution of alanine for Glu-378 and Glu-380 caused a complete loss of NADase and ARTase. However, exchange of Glu-378 to aspartic acid or glutamine resulted in little effect on NADase activity but a drastic reduction in ARTase activity (<0.1% of the wild-type activity). Exchange of Glu-380 to aspartic acid caused a drastic reduction in NADase and ARTase activities (<0.1% of the wild-type activities) but did not completely diminish them; that of the residue to glutamine caused a complete loss of ARTase activity. Replacement of Ser-338 with alanine resulted in 0.7 to 2.3% wild-type activities, and that of Ser-340 and Thr-339 caused a reduction in these activities of 5 to 30% wild-type activities. The kinetic analysis showed that Arg-295 and Ser-338 also play an important role in the binding of NAD+ to ia, that Arg-295, Glu-380, and Ser-338 play a crucial role in the catalytic rate of NADase activity, and that these three amino acid residues and Glu-378 are essential for ARTase activity. The effect of amino acid replacement in ia on ARTase activity was similar to that on lethal and cytotoxic activities, suggesting that lethal and cytotoxic activities in ia are dependent on ARTase activity.

* Corresponding author. Mailing address: Department of Microbiology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Yamashiro-cho, Tokushima 770-8514, Japan. Phone: 81 088-622-9611. Fax: 81 088-655-3051. E-mail: sakurai{at}ph.bunri-u.ac.jp.

Journal of Bacteriology, April 2000, p. 2096-2103, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Sundriyal, A., Roberts, A. K., Shone, C. C., Acharya, K. R. (2009). Structural Basis for Substrate Recognition in the Enzymatic Component of ADP-ribosyltransferase Toxin CDTa from Clostridium difficile. J. Biol. Chem. 284: 28713-28719 [Abstract] [Full Text]  
  • Tsuge, H., Nagahama, M., Oda, M., Iwamoto, S., Utsunomiya, H., Marquez, V. E., Katunuma, N., Nishizawa, M., Sakurai, J. (2008). Structural basis of actin recognition and arginine ADP-ribosylation by Clostridium perfringens {iota}-toxin. Proc. Natl. Acad. Sci. USA 105: 7399-7404 [Abstract] [Full Text]  
  • Fehr, D., Burr, S. E., Gibert, M., d'Alayer, J., Frey, J., Popoff, M. R. (2007). Aeromonas Exoenzyme T of Aeromonas salmonicida Is a Bifunctional Protein That Targets the Host Cytoskeleton. J. Biol. Chem. 282: 28843-28852 [Abstract] [Full Text]  
  • O'Neal, C. J., Jobling, M. G., Holmes, R. K., Hol, W. G. J. (2005). Structural Basis for the Activation of Cholera Toxin by Human ARF6-GTP. Science 309: 1093-1096 [Abstract] [Full Text]  
  • Barth, H., Aktories, K., Popoff, M. R., Stiles, B. G. (2004). Binary Bacterial Toxins: Biochemistry, Biology, and Applications of Common Clostridium and Bacillus Proteins. Microbiol. Mol. Biol. Rev. 68: 373-402 [Abstract] [Full Text]  
  • Nagahama, M., Yamaguchi, A., Hagiyama, T., Ohkubo, N., Kobayashi, K., Sakurai, J. (2004). Binding and Internalization of Clostridium perfringens Iota-Toxin in Lipid Rafts. Infect. Immun. 72: 3267-3275 [Abstract] [Full Text]  
  • Evans, H. R., Sutton, J. M., Holloway, D. E., Ayriss, J., Shone, C. C., Acharya, K. R. (2003). The Crystal Structure of C3stau2 from Staphylococcus aureus and Its Complex with NAD. J. Biol. Chem. 278: 45924-45930 [Abstract] [Full Text]  
  • Nagahama, M., Nagayasu, K., Kobayashi, K., Sakurai, J. (2002). Binding Component of Clostridium perfringens Iota-Toxin Induces Endocytosis in Vero Cells. Infect. Immun. 70: 1909-1914 [Abstract] [Full Text]  
  • Knapp, O., Benz, R., Gibert, M., Marvaud, J. C., Popoff, M. R. (2002). 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. 277: 6143-6152 [Abstract] [Full Text]  
  • Marvaud, J.-C., Smith, T., Hale, M. L., Popoff, M. R., Smith, L. A., Stiles, B. G. (2001). Clostridium perfringens Iota-Toxin: Mapping of Receptor Binding and Ia Docking Domains on Ib. Infect. Immun. 69: 2435-2441 [Abstract] [Full Text]  
  • Stabb, E. V., Reich, K. A., Ruby, E. G. (2001). Vibrio fischeri Genes hvnA and hvnB Encode Secreted NAD+-Glycohydrolases. J. Bacteriol. 183: 309-317 [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»