Immunochemical identification of the ADP-ribosyltransferase in botulinum C1 neurotoxin as C3 exoenzyme - like molecule

Narito Morii, Yasuhiro Ohashi, Yasuo Nemoto, Motohatsu Fujiwara, Yasuchika Ohnishi, Teiichi Nishiki, Yoichi Kamata, Shunji Kozaki, Shuh Narumiya, Genji Sakaguchi

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Abstract

Botulinum C1 neurotoxin and C3 exoenzyme were purified to apparent homogeneity from the culture filtrate of Clostridium botulinum type C strain 003-9. Both preparations catalyzed ADP-ribosylation of the same substrate, the Mr 22, 000 rho gene product (Gb). When the light and heavy chains of C1 toxin were separated, ADP-ribosyltransferase activity in the toxin was quantitatively recovered in the light chain fraction. Anti-C1 toxin antiserum precipitated the ADP-ribosyltransferase activity and the neurotoxicity of C1 toxin in parallel, whereas it had no effect on C3 exoenzyme. On the other hand, anti-C3 exoenzyme anti serum precipitated the ADP-ribosyltransferase activities of both C3 exoenzyme and C1 toxin. This antibody, however, did not precipitate the neurotoxicity of C1 toxin. The ADP-ribosyltransferase in C1 toxin was quantitatively adsorbed onto the anti-C3 antibody column and separated from the majority of C1 toxin protein. The enzyme was then eluted with acidic urea and Western blotting analysis of this eluate revealed the appearance of a protein band positively stained with anti-C3 antibody at a position similar to that of C3 exoenzyme. Quantitative determination by enzyme-linked immunosorbent assay showed that the C3-like immunoreactivity is present in the C1 toxin molecules at the molecular ratio of 1 to 1, 000. These results suggest that the ADP-ribosyltransferase activity in Cl toxin is expressed by a C3-like molecule which is present in a small amount in the toxin preparation and appears to bind to the toxin components). The above results also indicate that the ADP-ribosyltransferase in C1 toxin is not related to its neurotoxin action.

Original languageEnglish
Pages (from-to)769-775
Number of pages7
JournalJournal of biochemistry
Volume107
Issue number5
DOIs
Publication statusPublished - May 1990

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Morii, N., Ohashi, Y., Nemoto, Y., Fujiwara, M., Ohnishi, Y., Nishiki, T., Kamata, Y., Kozaki, S., Narumiya, S., & Sakaguchi, G. (1990). Immunochemical identification of the ADP-ribosyltransferase in botulinum C1 neurotoxin as C3 exoenzyme - like molecule. Journal of biochemistry, 107(5), 769-775. https://doi.org/10.1093/oxfordjournals.jbchem.a123123