1887

Abstract

Summary: Two types of subunit, with molecular weights estimated to be 28000 and 8000, were demonstrated in the cholera exo-enterotoxin by sodium dodecylsulphate polyacrylamide electrophoresis. The light (L) component but not the heavy (H) was demonstrable in the antigenically identical natural toxoid. A spilt of the H component by reduction and alkylation suggested it to be a disulphide-bridged dimer.

Gel filtration at acid pH permitted separation of the H and L subunits in isolated form. Fractions with isolated subunits were non-toxic (skin test), in contrast to fractions with intermediate filtration rate containing both subunits. Fractions with a L:H ratio of about 3.5 were even more toxic than the intact toxin (L:H ratio about 2). Addition of the isolated subunits to toxic fractions generally caused inhibition and only exceptionally potentiation of toxicity.

The data suggest that the cholera toxin might consist of an aggregate of seven L and one H subunits while the natural toxoid seems composed of only the aggregated L chains.

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/content/journal/micro/10.1099/00221287-76-2-417
1973-06-01
2021-10-23
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References

  1. Craig J. P. 1965; A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera. Nature, London 207:614–616
    [Google Scholar]
  2. Craig J. P. 1972; The enterotoxic enteropathies. In Microbial Pathogenicity in Man and Animals pp 129–155 Edited by Smith H., Pearce J. H. Cambridge University Press;
    [Google Scholar]
  3. Dunker A. K., Rueckert R. R. 1969; Observations on molecular weight determinations on polyacrylamide gel. Journal of Biological Chemistry 244:5074–5080
    [Google Scholar]
  4. Finkelstein R. A., Lo Spalluto J. J. 1969; Pathogenesis of experimental cholera. Preparation and isolation of choleragen and choleragenoid. Journal of Experimental Medicine 130:185–202
    [Google Scholar]
  5. Finkelstein R. A., Lo Spalluto J. J. 1970; Production of highly purified choleragen and choleragenoid. Journal of Infectious Diseases 121:63–72
    [Google Scholar]
  6. Hanson L. Å., Holmgren J., Wadsworth C. 1971; A radial immuno-gel filtration method for characterization and quantitation of macromolecules. International Archives of Allergy and Applied Immunology 40:806–819
    [Google Scholar]
  7. van Heyningen W. E. 1970; General characteristics (of bacterial protein toxins). In Microbial Toxins vol 1 pp 1–28 Edited by Ajl S. J., Kadis S., Montie T. C. New York and London: Academic Press;
    [Google Scholar]
  8. Holmgren J., Lönnroth I., Ouchterlony Ö. 1971; Immunochemical studies of two cholera toxin-containing preparations of Vibrio cholerae . Infection and Immunity 3:747–755
    [Google Scholar]
  9. Holmgren J., Lönnroth I., Ouchterlony Ö., Svennerholm A.-M. 1972; Studies on cholera toxins. Journal of General Microbiology 73:xxix
    [Google Scholar]
  10. Holmgren J., Lönnroth I., Svennerholm L. 1973; Fixation and inactivation of cholera toxin by GM1 ganglioside. Scandinavian Journal of Infectious Diseases 5:77–78
    [Google Scholar]
  11. Kabat E. A., Mayer M. M. 1961 Experimental Immunochemistry, 2nd edn.. pp 458–461 Springfield, Illinois: Charles C. Thomas;
    [Google Scholar]
  12. Laurell C.-B. 1966; Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Analytical Biochemistry 15:45–52
    [Google Scholar]
  13. LoSpalluto J. J., Finkelstein R. A. 1972; Chemical and physical properties of cholera exo-enterotoxin (choleragen) and its spontaneously formed toxoid (choleragenoid). Biochimica et biophysica acta 257:158–166
    [Google Scholar]
  14. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1966; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  15. Pierce N. F., Greenough W. B. III, Carpenter C. C. J. 1971; Vibrio cholerae enterotoxin and its mode of action. Bacteriological Reviews 35:1–13
    [Google Scholar]
  16. Wadsworth C. 1957; A slide microtechnique for the analysis of immune precipitates in gel. International Archives of Allergy and Applied Immunology 10:355–360
    [Google Scholar]
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