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. 1981 Mar;41(3):675–678. doi: 10.1128/aem.41.3.675-678.1981

Nature of intracellular type A botulinum neurotoxin.

E P Krysinski, H Sugiyama
PMCID: PMC243758  PMID: 7013709

Abstract

The neurotoxin in cells of young Clostridium botulinum type A culture was extracted with lysozyme. Highly purified neurotoxin preparation, obtained by processing the extract in two chromatographic steps had only unnicked (single-chain) molecules of molecular weight comparable to that of the dichains isolated from type A crystals. Trypsinization converted the unnicked molecules into dichains whose component subunits were of sizes indistinguishable from those of the neurotoxin from crystals. The enzymatic treatment increased toxicity of crude extract 30-fold but did not activate the purified intracellular neurotoxin preparation. The results indicated that intracellular type A botulinum neurotoxin is unnicked, is not fully activated, and is activated in the time between its extraction and purification. Since trypsinization nicked all of the single chains without increasing toxicity, nicking was not causally related to toxicity activation.

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Selected References

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

  1. BONVENTRE P. F., KEMPE L. L. Physiology of toxin production by Clostridium botulinum types A and B. I. Growth, autolysis, and toxin production. J Bacteriol. 1960 Jan;79:18–23. doi: 10.1128/jb.79.1.18-23.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. DasGupta B. R., Sugiyama H. A common subunit structure in Clostridium botulinum type A, B and E toxins. Biochem Biophys Res Commun. 1972 Jul 11;48(1):108–112. doi: 10.1016/0006-291x(72)90350-6. [DOI] [PubMed] [Google Scholar]
  3. DasGupta B. R., Sugiyama H. Role of arginine residues in the structure and biological activity of botulinum neurotoxin types A and E. Biochem Biophys Res Commun. 1980 Mar 28;93(2):369–375. doi: 10.1016/0006-291x(80)91086-4. [DOI] [PubMed] [Google Scholar]
  4. DasGupta B. R., Sugiyama H. Single chain and dichain forms of neurotoxin in type F Clostridium botulinum culture. Toxicon. 1977;15(5):466–471. doi: 10.1016/0041-0101(77)90128-3. [DOI] [PubMed] [Google Scholar]
  5. Dasgupta B. R., Sugiyama H. Isolation and characterization of a protease from Clostridium botulinum type B. Biochim Biophys Acta. 1972 Jun 16;268(3):719–729. doi: 10.1016/0005-2744(72)90276-8. [DOI] [PubMed] [Google Scholar]
  6. Dasgupta B. R., Sugiyama H. Molecular forms of neurotoxins in proteolytic Clostridium botulinum type B cultures. Infect Immun. 1976 Sep;14(3):680–686. doi: 10.1128/iai.14.3.680-686.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Knox J., Brown W. P., Spero L. The role of sulfhydryl groups in the activity of type A botulinum toxin. Biochim Biophys Acta. 1970 Aug 21;214(2):350–354. doi: 10.1016/0005-2795(70)90012-7. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Miura T. Isolation and characterization of an esterase-active enzyme from pronase with special reference to activation of Clostridium botulinum type E progenitor toxin. Jpn J Med Sci Biol. 1974 Dec;27(6):285–296. doi: 10.7883/yoken1952.27.285. [DOI] [PubMed] [Google Scholar]
  10. Miyazaki S., Iwasaki M., Sakaguchi G. Clostridium botulinum type D toxin: purification, molecular structure, and some immunological properties. Infect Immun. 1977 Aug;17(2):395–401. doi: 10.1128/iai.17.2.395-401.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Moberg L. J., Sugiyama H. Affinity chromatography purification of type A botulinum neurotoxin from crystalline toxic complex. Appl Environ Microbiol. 1978 May;35(5):878–880. doi: 10.1128/aem.35.5.878-880.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Ohishi I., Sakaguchi G. Response of type B and E Botulinum toxins to purified sulfhydryl-dependent protease produced by Clostridium botulinum type F. Jpn J Med Sci Biol. 1977 Aug;30(4):179–190. doi: 10.7883/yoken1952.30.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Sugiyama H., Moberg L. J., Messer S. L. Improved procedure for crystallization of Clostridium botulinum type A toxic complexes. Appl Environ Microbiol. 1977 Apr;33(4):963–966. doi: 10.1128/aem.33.4.963-966.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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