1887

Abstract

Lethal toxin from composed of protective antigen (PA) and lethal factor (LF). Anti-PA mAbs that neutralized lethal toxin activity, either or , identified three non-overlapping antigenic regions on PA. Two distinct antigenic regions were recognized by the four mAbs that neutralized lethal toxin activity by inhibiting the binding of I-LF to cell-bound PA. Mapping showed that one mAb, 1G3, recognized an epitope on a 17 IcDa fragment located between amino acid residues Ser-168 and Phe-314. The three other mAbs, 2D3, 2D5 and 10D2, recognized an epitope between amino acids Ile-581 and Asn-601. A single antigenic region was recognized by the three mAbs, 3B6, 14B7 and 10E10, that inhibited binding of I-PA to cells. This region was located between amino acids Asp-671 and lle-721. These results confirm previously defined functional domains of PA and suggest that LF may interact with two different sites on PA to form lethal toxin.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-3-707
1996-03-01
2021-05-08
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/3/mic-142-3-707.html?itemId=/content/journal/micro/10.1099/13500872-142-3-707&mimeType=html&fmt=ahah

References

  1. Beall F. A., Taylor M. J., Thorne C. B. 1962; Rapid lethal effect in rats of a third component found upon fractionating the toxin of Bacillus anthracis . J Bacterial 83:1274–1280
    [Google Scholar]
  2. Bhatnagar R., Singh Y., Leppla S. H., Friedlander A. M. 1989; Calcium is required for the expression of anthrax lethal toxin activity in the macrophage-like cell line J774A.1. Infect Immun 57:2107–2114
    [Google Scholar]
  3. Brennand D. M., Danson M. J., Hough D. W. 1986; A comparison of ELISA screening methods for the production of monoclonal antibodies against soluble protein antigens. J Immunol Methods 93:9–14
    [Google Scholar]
  4. Earley E. M., Osterling M. C. 1985; Fusion of mouse-mouse cells to produce hybridoma secreting monoclonal antibody. J Tissue Cult Methods 9:141–146
    [Google Scholar]
  5. Escuyer V., Collier R. J. 1991; Anthrax protective antigen interacts with a specific receptor on the surface of CHO-Kl cells. Infect Immun 59:3381–3386
    [Google Scholar]
  6. Ezzell J. W., Ivins B. E., Leppla S. H. 1984; Immuno- electrophoretic analysis, toxicity, and kinetics of in vitro production of the protective antigen and lethal factor components of Bacillus anthracis toxin. Infect Immun 45:761–767
    [Google Scholar]
  7. Fish D. C., Mahlandt B. G., Dobbs J. P., Lincoln R. E. 1968; Purification and properties of in vitro-produced anthrax toxin components. J Bacterial 95:907–918
    [Google Scholar]
  8. Friedlander A. M. 1986; Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem 261:7123–7126
    [Google Scholar]
  9. Gordon V. M., Leppla S. H., Hewlett E. L. 1988; Inhibitors of receptor-mediated endocytosis block the entry of Bacillus anthracis adenylate cyclase toxin but not that of Bordetella pertussis adenylate cyclase toxin. Infect Immun 56:1066–1069
    [Google Scholar]
  10. Green L. M., Reade J. L., Ware C. F. 1984; Rapid colorimetric assay for cell viability : application to the quantitation of cytotoxic and growth inhibitory lymphokines. J Immunol Methods 70:257–268
    [Google Scholar]
  11. Hollander Z., Katchalski-Katzir E. 1986; Use of monoclonal antibodies to detect conformational alterations in lactate dehydrogenase isoenzyme 5 on heat denaturation and on adsorption to polystyrene plates. Mol Immunol 23:927–933
    [Google Scholar]
  12. Klimpel K. R., Arora N., Leppla S. H. 1994; Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity. Mol Microbiol 13:1093–1100
    [Google Scholar]
  13. Kochi S. K., Schiavo G., Mock M., Montecucco C. 1994; Zinc content of the Bacillus anthracis lethal factor. FEMS Microbiol Lett 124:343–348
    [Google Scholar]
  14. Leppla S. H. 1982; Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc Natl Acad Sci USA 793162–3166
    [Google Scholar]
  15. Leppla S. H. 1988; Production and purification of anthrax toxin. Methods Enzymol 165:103–116
    [Google Scholar]
  16. Leppla S. H. 1991; The anthrax toxin complex. In Sourcebook of Bacterial Protein Toxins277–302 Alouf J. E., Freer J. H. New York: Academic Press;
    [Google Scholar]
  17. Leppla S. H., Friedlander A. M., Cora E. M. 1987; Proteolytic activation of anthrax toxin bound to cellular receptors. In Bacterial Protein Toxins111–112 Fehrenbach F., Alouf J. E., Falmagne P., Goebel W., Jeljaszewicz J., Jurgens D., Rappuoli R. Stuttgart: Gustav Fischer;
    [Google Scholar]
  18. Leppla S. H., Friedlander A. M., Singh Y., Cora E. M., Bhatnagar R. 1990; A model for anthrax toxic action at the cellular level. Salisbury Med Bull 68:41–43
    [Google Scholar]
  19. Little S. F., Lowe J. R. 1991; Location of receptor-binding region of protective antigen from Bacillus anthracis . Biochem Biophys Res Commun 180:531–537
    [Google Scholar]
  20. Little S.F., Leppla S. H., Cora E. 1988; Production and characterization of monoclonal antibodies to the protective antigen component of Bacillus anthracis toxin. Infect Immun 56:1807–1813
    [Google Scholar]
  21. Little S. F., Leppla S. H., Friedlander A. M. 1990; Production and characterization of monoclonal antibodies against the lethal factor component of Bacillus anthracis lethal toxin. Infect Immun 58:1606–1613
    [Google Scholar]
  22. Milne J. C., Furlong D., Hanna P. C., Wall J. S., Collier R. J. 1994; Anthrax protective antigen forms oligomers during intoxication of mammalian cells. J Biol Chem 269:20607–20612
    [Google Scholar]
  23. Mosmann T. 1983; Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63
    [Google Scholar]
  24. Novak J. M., Stein M.-P., Little S. F., Leppla S. H., Friedlander A. M. 1992; Functional characterization of protease-treated Bacillus anthracis protective antigen. J Biol Chem 267:17186–17193
    [Google Scholar]
  25. Singh Y., Chaudhary V. K., Leppla S. H. 1989; A deleted variant of Bacillus anthracis protective antigen is non-toxic and blocks anthrax toxin action in vivo . J Biol Chem 264:19103–19107
    [Google Scholar]
  26. Singh Y., Klimpel K. R., Quinn C. P., Chaudhary V. K., Leppla S. H. 1991; The carboxyl-terminal end of protective antigen is required for receptor binding and anthrax toxin activity. J Biol Chem 266:15493–15497
    [Google Scholar]
  27. Singh Y., Klimpel K. R., Arora N, Sharma M., Leppla S. H. 1994; The chymotrypsin-sensitive site, FFD315, in anthrax toxin protective antigen is required for translocation of lethal factor. J Biol Chem 269:29039–29046
    [Google Scholar]
  28. Stanley J. L, Smith H. 1961; Purification of factor I and recognition of a third factor of the anthrax toxin. J Gen Microbiol 26:49–66
    [Google Scholar]
  29. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets : procedure and some applications. Proc Natl Acad Sci USA 764350–4354
    [Google Scholar]
  30. Welkos S. L., Lowe J. R., Eden-McCutchan F., Vodkin M., Leppla S. H., Schmidt J. J. 1988; Sequence and analysis of the DNA encoding protective antigen of Bacillus anthracis . Gene 69:287–300
    [Google Scholar]
  31. Wu X. C., Lee W., Tran L., Wong S. L. 1991; Engineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases. J Bacterial 173:4952–4958
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-3-707
Loading
/content/journal/micro/10.1099/13500872-142-3-707
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error