1887

Abstract

SUMMARY Factor I of the anthrax toxin was isolated and showed one major component in the ultracentrifuge and on paper electrophoresis; it contained less 5.5% of extraneous antigens detectable by serological precipitation in gels. The final preparation contained all the usual amino acids (N = 10.1%) and some carbohydrate (6%, calculated as glucose) and phosphorus (0.7%). The most striking aspects of its analysis were a high ash (10-13%) and a light absorption at 260 mμ. The high ash was not due to one element but to a highly variable metal content (mainly Ca, Mg, Ni, Cu) indicating a powerful and indiscriminate chelating action of factor I. This chelating action might have been due to the chemical entity which absorbed light at 260 mμ and which was not RNA or DNA.

The final preparation of factor I was not toxic when injected alone but when mixed with purified factor II it evoked oedema in the skin of a rabbit and killed mice. However, the concentrationof this mixture which killed mice formed a much larger skin reaction in rabbits than a comparable dose (based on mouse LD 50) of either curde toxin or a mixture of crude factors I and II. An investigationof this fact led toteh demonstraion and partial purification of a third factor (III) of the anthrax toxin which: (1) was different serologically from factors I and II; (2) was present in anthrax toxin produced (3) was non-toxic when injected alone; (4) was lethal for mice when mixed with factor II but not with factor I; (5) increased the lethality of mixtures of factors I and II for mice and decreased their capacity to produce oedema in the skin of rabbits. A mixture of factors I, II and III showed synergic action in toxicity tests in mice; the mixture killed guinea pigs which showed signs of ligaemic secondary shock (as did guinea pigs killed by anthrax infection).

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1961-09-01
2022-01-19
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References

  1. Fiske C. H., SubbaRow Y. 1925; The colorimetric determination of phosphorus. J. biol. Chem 66:375
    [Google Scholar]
  2. Folch J., Lees M., Sloane-Stanley G. H. 1954; A simple method for preparation of total pure lipid extracts from brain. Fed. Proc 13:209
    [Google Scholar]
  3. Harris-Smith P. W., Smith H., Keppie J. 1958; Production in vitro of the toxin of Bacillus anthracis previously recognized in vivo. J. gen. Microbiol 19:91
    [Google Scholar]
  4. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. J. Biol. Chem 193:265
    [Google Scholar]
  5. Peterson E. A., Sober H. A. 1956; Chromatography of proteins. I. Cellulose ionexchange adsorbents J. Amer. chem. Soc 78:751
    [Google Scholar]
  6. Sargeant K., Stanley J. L., Smith H. 1960; The serological relationship between purified preparations of factors I and II of the anthrax toxin produced in vivo and in vitro. J. gen. Microbiol 22:219
    [Google Scholar]
  7. Smith H., Gallop R. C. 1956; The chemical basis of the virulence of Bacillus anthracis. VI. An extracellular immunising aggressin isolated from exudates of infected guinea-pigs Brit. J. exp. Path 37:144
    [Google Scholar]
  8. Smith H., Keppie J., Stanley J. L. 1955a; The chemical basis of the virulence of Bacillus anthracis. V. The specific toxin produced by B. anthracis in vivo Brit. J. exp. Path 36:460
    [Google Scholar]
  9. Smith H., Keppie J., Stanley J. L., Harris-Smith P. W. 1955b; The chemical basis of the virulence of Bacillus anthracis. IV. Secondary shock as the major factor in death of guinea pigs from anthrax Brit. J. exp. Path 36:323 R G. Microb. xxvi
    [Google Scholar]
  10. Smith H., Tempest D. W., Stanley J. L., Harris-Smith P. W., Gallop R. C. 1956; The chemical basis of the virulence of Bacillus anthracis. VII. Two components of the anthrax toxin: Their relationship to known immunizing aggressins Brit. J. exp. Path 37:263
    [Google Scholar]
  11. Sørensen M., Haugaard G. 1933; Über die Anwendbarkeit der Orcinreaktion zur Bestimmung der Art und Menge von Kohlen-hydratgruppen in Eiweisst Stoffen. Biochem. Z 260:247
    [Google Scholar]
  12. Stanley J. L., Sargeant K., Smith H. 1960; Purification of factors I and II of the anthrax toxin produced in vivo. J. gen. Microbiol 22:206
    [Google Scholar]
  13. Strange R. E., Thorne C. B. 1958; Further purification studies on the protective antigen of Bacillus anthracis produced in vitro. J. Bact 76:192
    [Google Scholar]
  14. Thorne C. B., Belton F. C. 1957; An agar diffusion method for titrating Bacillus anthracis immunizing antigen and its application to a study of antigen production. J. gen. Microbiol 17:505
    [Google Scholar]
  15. Thorne C. B., Molnar D. M., Strange R. E. 1960; Production of toxin in vitro by Bacillus anthracis and its separation into two components. J. Bact 79:250
    [Google Scholar]
  16. Volkin E., Cohn W. E. 1954; Estimation of nucleic acids. In Methods of Biochemical Analysis ed Glick D. 1287 New York: Interscience Publishers Inc;
    [Google Scholar]
  17. Wade H. E. 1961; The auto-degradation of ribonucleoprotein in Escherichia coli. Biochem. J 78:457
    [Google Scholar]
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