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

 Previous Article  |  Next Article 

Infect. Immun., 05 1996, 1589-1594, Vol 64, No. 5
Copyright © 1996, American Society for Microbiology

Molecular composition of Clostridium botulinum type A progenitor toxins

K Inoue, Y Fujinaga, T Watanabe, T Ohyama, K Takeshi, K Moriishi, H Nakajima, K Inoue and K Oguma
Department of Bacteriology, Okayama University Medical School, Japan.

The molecular composition of progenitor toxins produced by a Clostridium botulinum type A strain (A-NIH) was analyzed. The strain produced three types of progenitor toxins (19 S, 16 S, and 12 S) as reported previously. Purified 19 S and 16 S toxins demonstrated the same banding profiles on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), indicating that they consist of the same protein components. The nontoxic components of the 19 S and 16 S toxins are a nontoxic non-hemagglutinin (HA) (molecular mass, 120 kDa) and HA. HA could be fractionated into five subcomponents with molecular masses of 52, 35, 20, 19, and 15 kDa in the presence of 2-mercaptoethanol. The molar ratios of neurotoxins, nontoxic non-HAs, and each HA subcomponent of the 19 S and 16 S toxins showed that only HA-35 of the 19 S toxin was approximately twice the size of that of the 16 S toxin, suggesting that the 19 S toxin is a dimer of the 16 S toxin cross-linked by the 35- kDa subcomponent. The nontoxic non-HA of the 12 S toxin, but not those of the 19 S and 16 S toxins, demonstrated two bands with molecular masses of 106 and 13 kDa on SDS-PAGE with or without 2-mercaptoethanol. It was concluded from the N-terminal amino acid sequences that 106- and 13-kDa proteins were generated by a cleavage of whole nontoxic non-HA. This may explain why the 12 S and 16 S (and 19 S) toxins exist in the same culture. We also found that the HA and its 35-kDa subcomponent exist in a free state in the culture fluid along with three types of progenitor toxins.

This article has been cited by other articles:

• Jacobson, M. J., Lin, G., Raphael, B., Andreadis, J., Johnson, E. A. (2008). Analysis of Neurotoxin Cluster Genes in Clostridium botulinum Strains Producing Botulinum Neurotoxin Serotype A Subtypes. Appl. Environ. Microbiol. 74: 2778-2786 [Abstract] [Full Text]  
• Fujinaga, Y. (2006). Transport of Bacterial Toxins into Target Cells: Pathways Followed by Cholera Toxin and Botulinum Progenitor Toxin. J Biochem 140: 155-160 [Abstract] [Full Text]  
• Couesnon, A., Raffestin, S., Popoff, M. R. (2006). Expression of botulinum neurotoxins A and E, and associated non-toxin genes, during the transition phase and stability at high temperature: analysis by quantitative reverse transcription-PCR.. Microbiology 152: 759-770 [Abstract] [Full Text]  
• Mutoh, S., Suzuki, T., Hasegawa, K., Nakazawa, Y., Kouguchi, H., Sagane, Y., Niwa, K., Watanabe, T., Ohyama, T. (2005). Four molecules of the 33 kDa haemagglutinin component of the Clostridium botulinum serotype C and D toxin complexes are required to aggregate erythrocytes. Microbiology 151: 3847-3858 [Abstract] [Full Text]  
• Lee, J.-C., Yokota, K., Arimitsu, H., Hwang, H.-J., Sakaguchi, Y., Cui, J., Takeshi, K., Watanabe, T., Ohyama, T., Oguma, K. (2005). Production of anti-neurotoxin antibody is enhanced by two subcomponents, HA1 and HA3b, of Clostridium botulinum type B 16S toxin-haemagglutinin. Microbiology 151: 3739-3747 [Abstract] [Full Text]  
• Hines, H. B., Lebeda, F., Hale, M., Brueggemann, E. E. (2005). Characterization of Botulinum Progenitor Toxins by Mass Spectrometry. Appl. Environ. Microbiol. 71: 4478-4486 [Abstract] [Full Text]  
• Franciosa, G., Floridi, F., Maugliani, A., Aureli, P. (2004). Differentiation of the Gene Clusters Encoding Botulinum Neurotoxin Type A Complexes in Clostridium botulinum Type A, Ab, and A(B) Strains. Appl. Environ. Microbiol. 70: 7192-7199 [Abstract] [Full Text]  
• Maksymowych, A. B., Simpson, L. L. (2004). Structural Features of the Botulinum Neurotoxin Molecule That Govern Binding and Transcytosis across Polarized Human Intestinal Epithelial Cells. J. Pharmacol. Exp. Ther. 310: 633-641 [Abstract] [Full Text]  
• Arimitsu, H., Lee, J.-C., Sakaguchi, Y., Hayakawa, Y., Hayashi, M., Nakaura, M., Takai, H., Lin, S.-N., Mukamoto, M., Murphy, T., Oguma, K. (2004). Vaccination with Recombinant Whole Heavy Chain Fragments of Clostridium botulinum Type C and D Neurotoxins. CVI 11: 496-502 [Abstract] [Full Text]  
• Fujinaga, Y., Inoue, K., Watarai, S., Sakaguchi, Y., Arimitsu, H., Lee, J.-C., Jin, Y., Matsumura, T., Kabumoto, Y., Watanabe, T., Ohyama, T., Nishikawa, A., Oguma, K. (2004). Molecular characterization of binding subcomponents of Clostridium botulinum type C progenitor toxin for intestinal epithelial cells and erythrocytes. Microbiology 150: 1529-1538 [Abstract] [Full Text]  
• Inoue, K., Sobhany, M., Transue, T. R., Oguma, K., Pedersen, L. C., Negishi, M. (2003). Structural analysis by X-ray crystallography and calorimetry of a haemagglutinin component (HA1) of the progenitor toxin from Clostridium botulinum. Microbiology 149: 3361-3370 [Abstract] [Full Text]  
• Tamai, E., Ishida, T., Miyata, S., Matsushita, O., Suda, H., Kobayashi, S., Sonobe, H., Okabe, A. (2003). Accumulation of Clostridium perfringens Epsilon-Toxin in the Mouse Kidney and Its Possible Biological Significance. Infect. Immun. 71: 5371-5375 [Abstract] [Full Text]  
• Matsushima-Hibiya, Y., Watanabe, M., Hidari, K. I.-P. J., Miyamoto, D., Suzuki, Y., Kasama, T., Kanazawa, T., Koyama, K., Sugimura, T., Wakabayashi, K. (2003). Identification of Glycosphingolipid Receptors for Pierisin-1, a Guanine-specific ADP-ribosylating Toxin from the Cabbage Butterfly. J. Biol. Chem. 278: 9972-9978 [Abstract] [Full Text]  
• Arimitsu, H., Inoue, K., Sakaguchi, Y., Lee, J., Fujinaga, Y., Watanabe, T., Ohyama, T., Hirst, R., Oguma, K. (2003). Purification of Fully Activated Clostridium botulinum Serotype B Toxin for Treatment of Patients with Dystonia. Infect. Immun. 71: 1599-1603 [Abstract] [Full Text]  
• MAHMUT, N., INOUE, K., FUJINAGA, Y., ARIMITSU, H., SAKAGUCHI, Y., HUGHES, L., HIRST, R., MURPHY, T., TSUJI, T., WATANABE, T., OHYAMA, T., KARASAWA, T., NAKAMURA, S., YOKOTA, K., OGUMA, K. (2002). Mucosal immunisation with Clostridium botulinum type C 16 S toxoid and its non-toxic component. J Med Microbiol 51: 813-820 [Abstract] [Full Text]  
• Inoue, K., Fujinaga, Y., Honke, K., Arimitsu, H., Mahmut, N., Sakaguchi, Y., Ohyama, T., Watanabe, T., Inoue, K., Oguma, K. (2001). Clostridium botulinum type A haemagglutinin-positive progenitor toxin (HA+-PTX) binds to oligosaccharides containing Gal{beta}1-4GlcNAc through one subcomponent of haemagglutinin (HA1). Microbiology 147: 811-819 [Abstract] [Full Text]  
• Kanazawa, T., Watanabe, M., Matsushima-Hibiya, Y., Kono, T., Tanaka, N., Koyama, K., Sugimura, T., Wakabayashi, K. (2001). Distinct roles for the N- and C-terminal regions in the cytotoxicity of pierisin-1, a putative ADP-ribosylating toxin from cabbage butterfly, against mammalian cells. Proc. Natl. Acad. Sci. USA 98: 2226-2231 [Abstract] [Full Text]  
• Schiavo, G., Matteoli, M., Montecucco, C. (2000). Neurotoxins Affecting Neuroexocytosis. Physiol. Rev. 80: 717-766 [Abstract] [Full Text]  
• Watanabe, M., Kono, T., Matsushima-Hibiya, Y., Kanazawa, T., Nishisaka, N., Kishimoto, T., Koyama, K., Sugimura, T., Wakabayashi, K. (1999). Molecular cloning of an apoptosis-inducing protein, pierisin, from cabbage butterfly: Possible involvement of ADP-ribosylation in its activity. Proc. Natl. Acad. Sci. USA 96: 10608-10613 [Abstract] [Full Text]  
• Maksymowych, A. B., Reinhard, M., Malizio, C. J., Goodnough, M. C., Johnson, E. A., Simpson, L. L. (1999). Pure Botulinum Neurotoxin Is Absorbed from the Stomach and Small Intestine and Produces Peripheral Neuromuscular Blockade. Infect. Immun. 67: 4708-4712 [Abstract] [Full Text]  
• Chen, F., Kuziemko, G. M., Stevens, R. C. (1998). Biophysical Characterization of the Stability of the 150-Kilodalton Botulinum Toxin, the Nontoxic Component, and the 900-Kilodalton Botulinum Toxin Complex Species. Infect. Immun. 66: 2420-2425 [Abstract] [Full Text]  
• Kouguchi, H., Watanabe, T., Sagane, Y., Sunagawa, H., Ohyama, T. (2002). In Vitro Reconstitution of the Clostridium botulinum Type D Progenitor Toxin. J. Biol. Chem. 277: 2650-2656 [Abstract] [Full Text]