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. 1985 May;48(2):312–317. doi: 10.1128/iai.48.2.312-317.1985

Purification and characterization of neurotoxin produced by Clostridium botulinum type C 6813.

J Terajima, B Syuto, J O Ochanda, S Kubo
PMCID: PMC261306  PMID: 3988338

Abstract

The toxin produced by Clostridium botulinum type C 6813 (C-6813) was purified 1,009-fold from the culture supernatant in an overall yield of 30%. The specific toxicity was 1.1 X 10(7) mouse minimum lethal doses per mg of protein. The toxin had a molecular weight of 144,000, composed of the light and heavy chains with molecular weights of 52,000 and 92,000, respectively, linked by one or two disulfide bond(s). The purified C-6813 toxin heavy and light chains reacted strongly with anti-type D heavy chain immunoglobulin G and anti-type C1 light chain immunoglobulin G, respectively. The amino acid compositions of C-6813 toxin heavy and light chains were more similar to those of type D heavy chain and type C1 light chain than to those of type C1 heavy chain and type D light chain, respectively. These results suggest that in the toxin produced by the type C strain at least two subtypes exist.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baillie R. D., Horowitz P. M. Denaturation-induced disulfide formation in the enzyme rhodanese. Biochim Biophys Acta. 1976 Apr 8;429(2):383–390. doi: 10.1016/0005-2744(76)90286-2. [DOI] [PubMed] [Google Scholar]
  2. Boroff D. A., Fleck U. Statistical analysis of a rapid in vivo method for the titration of the toxin of Clostridium botulinum. J Bacteriol. 1966 Nov;92(5):1580–1581. doi: 10.1128/jb.92.5.1580-1581.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  4. Goodwin T. W., Morton R. A. The spectrophotometric determination of tyrosine and tryptophan in proteins. Biochem J. 1946;40(5-6):628–632. doi: 10.1042/bj0400628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. HIRS C. H. The oxidation of ribonuclease with performic acid. J Biol Chem. 1956 Apr;219(2):611–621. [PubMed] [Google Scholar]
  6. Inoue K., Iida H. Phage-conversion of toxigenicity in Clostridium botulinum types C and D. Jpn J Med Sci Biol. 1971 Feb;24(1):53–56. [PubMed] [Google Scholar]
  7. Iwasaki M., Ohishi I., Sakaguchi G. Evidence that botulinum C2 toxin has two dissimilar components. Infect Immun. 1980 Aug;29(2):390–394. doi: 10.1128/iai.29.2.390-394.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jansen B. C. The toxic antigenic factors produced by Clostridium botulinum types C and D. Onderstepoort J Vet Res. 1971 Jun;38(2):93–98. [PubMed] [Google Scholar]
  9. Jensen W. I., Duncan R. M. The susceptibility of the mallard duck (Anas platyrhynchos) to Clostridium botulinum C2 toxin. Jpn J Med Sci Biol. 1980 Apr;33(2):81–86. doi: 10.7883/yoken1952.33.81. [DOI] [PubMed] [Google Scholar]
  10. Kahn R., Rubin R. W. Quantitation of submicrogram amounts of protein using coomassie brilliant blue R on sodium dodecyl sulfate-polyacrylamide slab-gels. Anal Biochem. 1975 Jul;67(1):347–352. doi: 10.1016/0003-2697(75)90305-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Murayama S., Syuto B., Oguma K., Iida H., Kubo S. Comparison of Clostridium botulinum toxins type D and C1 in molecular property, antigenicity and binding ability to rat-brain synaptosomes. Eur J Biochem. 1984 Aug 1;142(3):487–492. doi: 10.1111/j.1432-1033.1984.tb08312.x. [DOI] [PubMed] [Google Scholar]
  13. Ochanda J. O., Syuto B., Oguma K., Iida H., Kubo S. Comparison of antigenicity of toxins produced by Clostridium botulinum type C and D strains. Appl Environ Microbiol. 1984 Jun;47(6):1319–1322. doi: 10.1128/aem.47.6.1319-1322.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Oguma K., Agui T., Syuto B., Kimura K., Iida H., Kubo S. Four different monoclonal antibodies against type C1 toxin of Clostridium botulinum. Infect Immun. 1982 Oct;38(1):14–20. doi: 10.1128/iai.38.1.14-20.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Oguma K., Murayama S., Syuto B., Iida H., Kubo S. Analysis of antigenicity of Clostridium botulinum type C1 and D toxins by polyclonal and monoclonal antibodies. Infect Immun. 1984 Feb;43(2):584–588. doi: 10.1128/iai.43.2.584-588.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Oguma K., Syuto B., Agui T., Iida H., Kubo S. Homogeneity and heterogeneity of toxins produced by Clostridium botulinum type C and D strains. Infect Immun. 1981 Nov;34(2):382–388. doi: 10.1128/iai.34.2.382-388.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oguma K., Syuto B., Iida H., Kubo S. Antigenic similarity of toxins produced by Clostridium botulinum type C and D strains. Infect Immun. 1980 Dec;30(3):656–660. doi: 10.1128/iai.30.3.656-660.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ohishi I., Iwasaki M., Sakaguchi G. Purification and characterization of two components of botulinum C2 toxin. Infect Immun. 1980 Dec;30(3):668–673. doi: 10.1128/iai.30.3.668-673.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ohishi I., Iwasaki M., Sakaguchi G. Vascular permeability activity of botulinum C2 toxin elicited by cooperation of two dissimilar protein components. Infect Immun. 1981 Mar;31(3):890–895. doi: 10.1128/iai.31.3.890-895.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sugiyama H. Clostridium botulinum neurotoxin. Microbiol Rev. 1980 Sep;44(3):419–448. doi: 10.1128/mr.44.3.419-448.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Syuto B., Kubo S. Isolation and molecular size of Clostridium botulinum type C toxin. Appl Environ Microbiol. 1977 Feb;33(2):400–405. doi: 10.1128/aem.33.2.400-405.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Syuto B., Kubo S. Separation and characterization of heavy and light chains from Clostridium botulinum type C toxin and their reconstitution. J Biol Chem. 1981 Apr 25;256(8):3712–3717. [PubMed] [Google Scholar]
  23. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]

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