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. 1977 Aug;17(2):395–401. doi: 10.1128/iai.17.2.395-401.1977

Clostridium botulinum type D toxin: purification, molecular structure, and some immunological properties.

S Miyazaki, M Iwasaki, G Sakaguchi
PMCID: PMC421134  PMID: 892912

Abstract

Clostridium botulinum type D progenitor toxin was purified. The addition of ribonucleic acid to the whole culture helped initial acid precipitation of the toxin. As with type B, both L (16S) and M toxins (12S) obtained from a hemagglutinin-positive strain, whereas M toxin only was produced by a hemagglutinin-negative strain. M toxin (molecular weight, 300,000) consisted of one molecule each of a toxic (molecular weight, 170,000) and a nontoxic component (molecular weight, 130,000); L toxin consisted of both components plus hemagglutinin. The specific toxicity of M toxin was 5 X 10(8) mean lethal doses per mg of N; that of L toxin was 2.4 X 10(8) mean lethal doses per mg of N. These toxins were fully or nearly fully active, but in un-nicked form. Trypsinization caused nicking in the toxic component, forming a molecule made up of two peptide chains with molecular weights of 110,000 and 60,000; there was little or no increase in toxicity. The toxic component of type D was not antigenically related to that of type C, whereas the nontoxic component was antigenically indistinguishable from that of type C. The toxicities of both L nad M toxins of the hemagglutinin-positive strain were increased twofold by trypsinization. Neither toxin contained the C2 toxic factor elaborated by C and D strain.

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Selected References

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  1. BOWMER E. J. PREPARATION AND ASSAY OF THE INTERNATIONAL STANDARDS FOR CLOSTRIDIUM BOTULINUM TYPES A, B, C, D AND E ANTITOXINS. Bull World Health Organ. 1963;29:701–709. [PMC free article] [PubMed] [Google Scholar]
  2. Beers W. H., Reich E. Isolation and characterization of Clostridium botulinum type B toxin. J Biol Chem. 1969 Aug 25;244(16):4473–4479. [PubMed] [Google Scholar]
  3. 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]
  4. CARDELLA M. A., DUFF J. T., WINGFIELD B. H., GOTTFRIED C. Studies on immunity to toxins of Clostridium botulinum. VI. Purification and detoxification of type D toxin and the immunological response to toxoid. J Bacteriol. 1960 Mar;79:372–378. doi: 10.1128/jb.79.3.372-378.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dasgupta B. R., Boroff D. A., Rothstein E. Chromatographic fractionation of the crystalline toxin of Clostridium botulinum type A. Biochem Biophys Res Commun. 1966 Mar 22;22(6):750–756. doi: 10.1016/0006-291x(66)90212-9. [DOI] [PubMed] [Google Scholar]
  6. Eklund M. W., Poysky F. T. Activation of a toxic component of Clostridium botulinum types C and D by trypsin. Appl Microbiol. 1972 Jul;24(1):108–113. doi: 10.1128/am.24.1.108-113.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eklund M. W., Poysky F. T. Interconversion of type C and D strains of Clostridium botulinum by specific bacteriophages. Appl Microbiol. 1974 Jan;27(1):251–258. doi: 10.1128/am.27.1.251-258.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Jansen B. C., Knoetze P. C. Tryptic activation of Clostridium botulinum type C beta toxin. Onderstepoort J Vet Res. 1971 Dec;38(4):237–238. [PubMed] [Google Scholar]
  10. 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]
  11. Kitamura M., Sakaguchi S., Sakaguchi G. Purification and some properties of Clostridium botulinum type-E toxin. Biochim Biophys Acta. 1968 Oct 21;168(2):207–217. doi: 10.1016/0005-2795(68)90144-x. [DOI] [PubMed] [Google Scholar]
  12. Kozaki S., Sakaguchi G. Antigenicites of fragments of Clostridium botulinum type B derivative toxin. Infect Immun. 1975 May;11(5):932–936. doi: 10.1128/iai.11.5.932-936.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kozaki S., Sakaguchi S., Sakaguchi G. Purification and some properties of progenitor toxins of Clostridium botulinum type B. Infect Immun. 1974 Oct;10(4):750–756. doi: 10.1128/iai.10.4.750-756.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kozaki S., Sugii S., Oishi I., Sakaguchi S., Sakaguchi G. Proceedings: Clostridium botulinum type A, B, E and F 12S toxins. Jpn J Med Sci Biol. 1975 Feb;28(1):70–72. [PubMed] [Google Scholar]
  15. 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]
  16. Ohishi I., Sakaguchi G. Activation of botulinum toxins in the absence of nicking. Infect Immun. 1977 Aug;17(2):402–407. doi: 10.1128/iai.17.2.402-407.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ohishi I., Sakaguchi G. Molecular construction of Clostridium botulinum type F progenitor toxin. Appl Microbiol. 1975 Apr;29(4):444–447. doi: 10.1128/am.29.4.444-447.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Oishi I., Sakaguchi G. Purification of Clostridium botuliunum type F progenitor toxin. Appl Microbiol. 1974 Dec;28(6):923–928. doi: 10.1128/am.28.6.923-928.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  20. STERNE M. Hemagglutination by Clostridium botulinum type D. Science. 1954 Apr 2;119(3092):440–441. doi: 10.1126/science.119.3092.440. [DOI] [PubMed] [Google Scholar]
  21. Sakaguchi G., Sakaguchi S. Development of antibody to each component of Clostridium botulinum type E progenitor toxin. Jpn J Med Sci Biol. 1973 Dec;26(5):187–195. doi: 10.7883/yoken1952.26.187. [DOI] [PubMed] [Google Scholar]
  22. Sakaguchi G., Sakaguchi S., Kozaki S., Sugii S., Oishi I. Cross reaction in reversed passive hemagglutination between Clostridium botulinum type A and B toxins and its avoidance by the sue of anti-toxic component immunoglobulin isolated by affinity chromatography. Jpn J Med Sci Biol. 1974 Jun;27(3):161–172. doi: 10.7883/yoken1952.27.161. [DOI] [PubMed] [Google Scholar]
  23. Shimizu T., Izumi N., Kondo H. Proceedings: Immunological specificities of Clostridium botulinum type C and D toxins. Jpn J Med Sci Biol. 1975 Oct-Dec;28(5-6):298–301. [PubMed] [Google Scholar]
  24. Sugii S., Sakaguchi G. Molecular construction of Clostridium botulinum type A toxins. Infect Immun. 1975 Dec;12(6):1262–1270. doi: 10.1128/iai.12.6.1262-1270.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. WENTZEL L. M., STERNE M., POLSON A. High toxicity of pure botulinum type D toxin. Nature. 1950 Oct 28;166(4226):739–740. doi: 10.1038/166739b0. [DOI] [PubMed] [Google Scholar]
  26. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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