The Anthrax Toxin

  • Carlo Petosa
  • Robert C. Liddington
Part of the Molecular Biology Intelligence Unit book series (MBIU)


Anthrax is a disease known since antiquity1 and one of the first bacterial infections whose etiology was definitively established. The disease is caused by the Gram-positive, aerobic, spore-forming Bacillus anthracis, first isolated in 1877 by Robert Koch.2 The study of anthrax led to the establishment of Koch’s postulates, a set of criteria for identifying an organism as the causative agent of a specific infection.2 Louis Pasteur’s use of heat-inactivated anthrax cultures to immunize against the disease is generally credited as the first instance of a bacterial vaccine.3 Anthrax is primarily a disease of herbivorous animals, particularly sheep and cattle.4 Humans may acquire the disease from infected animals, typically as a cutaneous infection characterized by black pustules5 (whence the naming of the disease after the Greek word for “coal”). The pulmonary infection known as wool-sorter’s disease results from the inhalation of anthrax spores, often as a result of handling contaminated raw wool, hides or animal hair, and can lead to death within days.6–8 Though increasingly rare in human populations, anthrax remains of interest for several reasons, including its continuing incidence in animal populations,9 interest in improving the efficacy of the human vaccine,4,10 the threat of its use as a weapon of biological warfare (see ref. 11), its potential applications in the development of new therapeutic strategies such as targeted toxins,12 and as an experimental system for studying molecular pathogenesis.13


Zinc Bacillus Oligomer Polypeptide Trypsin 


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  1. 1.
    Dirckx JH. Virgil on anthrax. Am J Dermatopathol 1981; 3: 191–95.PubMedCrossRefGoogle Scholar
  2. 2.
    Koch R. The aetiology of anthrax based on the ontogeny of the anthrax bacillus. Beitr Biol Pflanz 1877; 2: 277.Google Scholar
  3. 3.
    Pasteur L. De l’attenuation des virus et de leur retour a la virulence. CR Acad Sci 1881; 19: 429.Google Scholar
  4. 4.
    Hambleton P, Turnbull PCB. Anthrax vaccine development: a continuing story. In: Mizrahi A. Bacterial vaccines. Advances in Biotechnological Processes, Vol 13. New York: Alan R. Liss, 1990; 105–22.Google Scholar
  5. 5.
    Christie AB. Infectious diseases: epidemiology and clinical practice. Edition IV. Edinburgh: Churchill Livingstone, 1987; 983.Google Scholar
  6. 6.
    Bell JH. On anthrax and anthracaemia in wool sorters, heifers and sheep. Br Med J 1880; 2: 656.PubMedCrossRefGoogle Scholar
  7. 7.
    La Force FM. Woolsorters’ disease in England. NY Acad Med 1978; 54: 956.Google Scholar
  8. 8.
    Albrink WS, Brooks SM, Biron RE et al. Human inhalation anthrax: a report on three cases. Am J Pathol 1960; 36: 457.PubMedGoogle Scholar
  9. 9.
    FAO/OIE/Who. Animal Health Year Book. New York: Unipub, 1985.Google Scholar
  10. 10.
    Turnbull PC. Anthrax vaccines: past, present, and future. Vaccine 1991; 9: 533–39.PubMedCrossRefGoogle Scholar
  11. 11.
    Meselson M, Guillemin J, Hugh-Jones M et al. The Sverdlovsk anthrax outbreak of 1979. Science 1994; 266: 1202–28.PubMedCrossRefGoogle Scholar
  12. 12.
    Pastan I, Chaudhary V, Fitzgerald DJ et al. Recombinant toxins as novel therapeutic agents. Ann Rev Biochem 1992; 61: 331–54.PubMedCrossRefGoogle Scholar
  13. 13.
    Leppla SH. Anthrax toxins. In: Moss J, Iglewski B, Vaughan M, Tu AT, eds. Bacterial Toxins and Virulence Factors in Disease. New York: Marcel Dekker, 1995: 543–72.Google Scholar
  14. 14.
    Leppla SH. The anthrax toxin complex. In: Alouf JE, Freer JH, eds. Sourcebook of Bacterial Protein Toxins. San Diego: Academic Press, 1991; 277–302.Google Scholar
  15. 15.
    Mikesell P, Ivins BE, Ristroph JD et al. Evidence for plasmid-mediated toxin production in Bacillus anthracis. Infect Immun 1983; 39: 371–76.PubMedGoogle Scholar
  16. 16.
    Green BD, Battisti L, Koehler TM et al. Demonstration of a capsule plasmid in Bacillus anthracis. Infect Immun 1985; 49: 291–97.PubMedGoogle Scholar
  17. 17.
    Uchida I, Sekizaki T, Hashimoto K et al. Association of the encapsulation of Bacillus anthracis with a 60 megadalton plasmid. J Gen Microbiol 1985; 131: 363–67.PubMedGoogle Scholar
  18. 18.
    Uchida I, Hashimoto K, Terakado N. Virulence and immunogenicity in experimental animals of Bacillus anthracis strains harbouring or lacking 110 MDa and 60 MDa plasmids. J Gen Microbiol 1986; 132: 557–59.PubMedGoogle Scholar
  19. 19.
    Jriedlander AM. Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem 1986; 261: 7123–26.Google Scholar
  20. 20.
    Hambleton P, Carman JA, Melling J. Anthrax: the disease in relation to vaccines. Vaccine 1984; 2: 125–32.PubMedCrossRefGoogle Scholar
  21. 21.
    Stanley JL, Smith H. The three factors of anthrax toxin: their immunogenicity and lack of demonstrable enzymic activity. J Gen Microbiol 1963; 31: 329–37.PubMedCrossRefGoogle Scholar
  22. 22.
    Bragg TS, Robertson DL. Nucleotide sequence and analysis of the lethal factor gene (lef) from Bacillus anthracis. Gene 1989; 81: 45–54.PubMedCrossRefGoogle Scholar
  23. 23.
    Welkos SL, Lowe JR, Eden-McCutchan F et al. Sequence and analysis of the DNA encoding protective antigen of Bacillus anthracis. Gene 1988; 69: 287–300.PubMedCrossRefGoogle Scholar
  24. 24.
    Escuyer V, Duflot E, Sezer O et al. Structural homology between virulence-associated bacterial adenylate cyclases. Gene 1988; 71: 293–98.PubMedCrossRefGoogle Scholar
  25. 25.
    Robertson DL, Tippets MT, Leppla SH. Nucleotide sequence of the Bacillus anthracis edema factor gene (cya): a calmodulin-dependent adenylate cyclase. Gene 1988; 73: 363–71.PubMedCrossRefGoogle Scholar
  26. 26.
    Bartkus JM, Leppla SH. Transcriptional regulation of the protective antigen gene of Bacillus anthracis. Infect Immun 1989; 57: 2295–300.PubMedGoogle Scholar
  27. 27.
    Uchida I, Hornung JM, Thorne CB et al. Cloning and characterization of a gene whose product is a transactivator of anthrax toxin synthesis. J Bacteriol 1993; 175: 5329–38.PubMedGoogle Scholar
  28. 28.
    Cataldi A, Labruyère E, Mock M. Construction and characterization of antigen-deficient Bacillus anthracis strain. Mol Microbiol 1990; 4: 1111–17.PubMedCrossRefGoogle Scholar
  29. 29.
    Hornung JM, Thorne CB. Insertion mutations affecting pXO 1-associated toxin production in Bacillus anthracis. Abstr 91st Annu Meet Am Soc Microbiol 1991; p.98 Abstr D-121.Google Scholar
  30. 30.
    Pezard C, Berche P, Mock M. Contribution of individual toxin components to virulence of Bacillus anthracis. Infect Immun 1991; 59: 3472–77.PubMedGoogle Scholar
  31. 31.
    Fish DC, Klein F, Lincoln RE et al. Pathophysiological changes in the rat associated with anthrax toxin. J Infect Dis 118: 114–24.Google Scholar
  32. 32.
    Beall FA, Taylor MJ, Thorne CB. Rapid lethal effect in rats of a third component found upon fractionating the toxin of Bacillus anthracis. J Bacteriol 1962; 83: 1274–80.PubMedGoogle Scholar
  33. 33.
    Smith H, Stoner HB. Anthrax toxic complex. Fed Proc 1967; 26: 1554–57.PubMedGoogle Scholar
  34. 34.
    Li J. Bacterial toxins. Curr Opin Struct Biol 1992; 2: 545–56.CrossRefGoogle Scholar
  35. 35.
    Considine R, Simpson L. Cellular and molecular action of binary toxins possessing ADP-ribosyltransferase activity. Toxicon 1991; 29: 913–36.PubMedCrossRefGoogle Scholar
  36. 36.
    Friedlander AM. The anthrax toxins. In: Saelinger C, ed. Trafficking of Bacterial Toxins. CRC Press: Boca Raton, 1990: 121–28.Google Scholar
  37. 37.
    Leppla SH. Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations in eukaryotic cells. Proc Natl Acad Sci USA 1982; 79: 3162–66.PubMedCrossRefGoogle Scholar
  38. 38.
    Pezard C, Duflot, E, Mock M. Construction of Bacillus anthracis mutant strains producing a a single toxin component. J Gen Microbiol 1993; 139: 2459–63.PubMedCrossRefGoogle Scholar
  39. 39.
    Stephen J. Anthrax toxin. Pharm Therap 1991; 12: 501–13.CrossRefGoogle Scholar
  40. 40.
    Hanna PC, Kochi, S, Collier RJ. Biochemical and physiological changes induced by anthrax lethal toxin in J774 macrophage-like cells. Mol Biol Cell 1992; 3: 1269–77.PubMedGoogle Scholar
  41. 41.
    Hanna PC, Kruskal BA, Ezekowitz RAB et al. Role of macrophage oxidative burst in the action of anthrax lethal toxin. Molecular Medicine 1994; 1: 7–18.PubMedGoogle Scholar
  42. 42.
    Hanna PC, Acosta D, Collier RJ. On the role of macrophages in anthrax. Proc Natl Acad Sci USA 1993; 90: 10198–201.PubMedCrossRefGoogle Scholar
  43. 43.
    Escuyer V, Collier RJ. Anthrax protective antigen interacts with a specific receptor on the surface of CHO-Ki cells. Infect Immun 1991; 59: 3381–86.PubMedGoogle Scholar
  44. 44.
    Kumpel KR, Molloy SS, Thomas G et al. Anthrax toxin protective antigen is activated by a cell surface protease with the sequence specificity and catalytic properties of furin. Proc Natl Acad Sci USA 1992; 89: 10277–81.CrossRefGoogle Scholar
  45. 45.
    Molloy SS, Bresnahan PA, Leppla SH et al. Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen. J Biol Chem 1992; 267: 16396–402.PubMedGoogle Scholar
  46. 46.
    Gordon VM, Kumpel KR, Arora N et al. Proteolytic activation of bacterial toxins by eukaryotic cells is performed by furin and by additional cellular proteases. Infect Immun 1995; 63: 82–87.PubMedGoogle Scholar
  47. 47.
    Leppla SH, Friedlander AM, Cora EM. Proteolytic activation of anthrax toxin bound to cellular receptors. In: Fehrenbach FJ, Alouf JE, Falmagne P, Goebel W, Jeljaszewicz J, Jurgen D, Rappuoli, R, ed. Bacterial Protein Toxins. New York: Gustav Fischer, 1988: 111–12.Google Scholar
  48. 48.
    Gordon VM, Leppla SH, Hewlett EL. 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 1988; 56: 1066–69.PubMedGoogle Scholar
  49. 49.
    Klimpel KR, Arora N, Leppla SH. Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity. Mol Microbiol 1994; 13: 1093–100.PubMedCrossRefGoogle Scholar
  50. 50.
    Milne JC, Furlong D, Hanna PC et al. Anthrax protective antigen forms oligomers during intoxication of mammalian cells. J Biol Chem 1994; 269: 20607–12.PubMedGoogle Scholar
  51. 51.
    Leppla SH. Bacillus anthracis calmodulin-dependent adenylate cyclase: chemical and enzymatic properties and interactions with eukaryotic cells. Adv Cyclic Nucleotide Protein Phosphorylation Res 1984; 17:189–98.Google Scholar
  52. 52.
    Labruyère E, Mock M, Ladant D et al. Characterization of ATP and calmodulin-binding properties of a truncated form of Bacillus anthracis adenylate cyclase. Biochem 1990; 29: 4922–28.CrossRefGoogle Scholar
  53. 53.
    Goyard S, Orlando C, Sabaier J-M et al. Identification of a common domain in calmodulin-activated eukaryotic and bacterial adenylate cyclases. Biochem 1989; 28: 1964–67.CrossRefGoogle Scholar
  54. 54.
    Xia Z, Storm DR. A-type ATP binding consensus sequences are critical for the catalytic activity of the calmodulin-sensitive adenylyl cyclase form Bacillus anthracis. J Biol Chem 1990; 265: 6517–20.PubMedGoogle Scholar
  55. 55.
    Munier H, Blanco FJ, Prêcheur B et al. Characterization of a synthetic calmodulin-binding peptide derived from Bacillus anthracis adenylate cyclase. J Biol Chem 1993; 268: 1695–701.PubMedGoogle Scholar
  56. 56.
    Little SF, Leppla SH, Burnett JW et al. Structure-function analysis of Bacillus anthracis edema factor by using monoclonal antibodies. Biochem Biophys Res Commun 1994; 199: 676–82.PubMedCrossRefGoogle Scholar
  57. 57.
    Arora N, Klimpel KR, Singh Y et al. Fusions of anthrax toxin lethal factor to the ADP-ribosylation domain of Pseudomonas Exotoxin A are potent cytotoxins which are translocated to the cytosol of mammalian cells. J Biol Chem 1992; 267: 15542–48.PubMedGoogle Scholar
  58. 58.
    Arora N, Leppla SH. Residues 1–254 of anthrax toxin lethal factor are sufficient to cause cellular uptake of fused polypeptides. J Biol Chem 1993; 268: 3334–41.PubMedGoogle Scholar
  59. 59.
    Arora N, Leppla SH. Fusions of anthrax toxin lethal factor with shiga toxin and diphtheria toxin enzymatic domains are toxic to mammalian cells. Infect Immun 1994; 62: 4955–61.PubMedGoogle Scholar
  60. 60.
    Milne JC, Blanke SR, Hanna PC et al. Protective antigen-binding domain of anthrax lethal factor mediates translocation of a heterologous protein fused to its amino-or carboxy-terminus. Mol Microbiology 1995; 15: 661–66.CrossRefGoogle Scholar
  61. 61.
    Quinn CP, Singh Y, Klimpel KR et al. Functional mapping of anthrax toxin lethal factor by in-frame insertion mutagenesis. J Biol Chem 1991; 266: 20124–30.PubMedGoogle Scholar
  62. 62.
    Kochi SK, Schiavo G, Mock M et al. Zinc content of the Bacillus anthracis lethal factor. FEMS Microbiol Lett 1994; 124: 343–48.PubMedCrossRefGoogle Scholar
  63. 63.
    Jongeneel CV, Bouvier J, Bairoch A. A unique signature identifies a family of zinc-dependent metallopeptidases. FEBS Lett 1989; 242: 211–14.PubMedCrossRefGoogle Scholar
  64. 64.
    Vallee BL, Auld DS. Zinc coordination, function, and structure of zinc enzymes and other proteins. Biochemistry 1990; 29: 5647–59.PubMedCrossRefGoogle Scholar
  65. 65.
    Schiavo G, Benfenati F, Poulain B et al. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature 1992; 359: 832–35.PubMedCrossRefGoogle Scholar
  66. 66.
    Montecucco C, Schiavo G. Tetanus and botulism neurotoxins: a new group of zinc proteases. Trend Biochem Sci 1993; 18: 324–27.PubMedCrossRefGoogle Scholar
  67. 67.
    Oguma K, Fujinaga Y, Inoue K. Structure and function of Clostridium botulinum toxins. Microbiol Immunol 1995; 39: 161–68.PubMedGoogle Scholar
  68. 68.
    Novak JM, Stein M-P, Little SF et al. Functional characterization of protease-treated Bacillus anthracis protective antigen. J Biol Chem 1992; 267: 17186–93.PubMedGoogle Scholar
  69. 69.
    Singh Y, Chaudhary VK, Leppla SH. A deleted variant of Bacillus anthracis protective antigen is nontoxic and blocks anthrax toxin in vivo. J Biol Chem 1989; 264: 19103–37.PubMedGoogle Scholar
  70. 70.
    Singh Y, Klimpel KR, Arora N et al. The chymotrypsin-sensitivie site, FFD31$, in anthrax toxin protective antigen is required for translocation of lethal factor. J Biol Chem 1994; 269: 29039–46.PubMedGoogle Scholar
  71. 71.
    Singh Y, Klimpel KR, Quinn CP et al. The carboxyl-terminal end of protective antigen is required for receptor binding and anthrax toxin activity. J Biol Chem 1991; 266: 15493–97.PubMedGoogle Scholar
  72. 72.
    Little SF, Lowe JR. Location of receptor-binding region of protective antigen from Bacillus anthracis. Biochem Biophys Res Commun 1991; 180: 531–37.PubMedCrossRefGoogle Scholar
  73. 73.
    Blaustein RO, Koehler TM, Collier RJ et al. Anthrax toxin: channel-forming activity of protective antigen in planar phospholipid bilayers. Proc Natl Acad Sci USA 1989; 86: 2209–13.PubMedCrossRefGoogle Scholar
  74. 74.
    Finkelstein A. The channel formed in planar lipid bilayers by the protective antigen component of anthrax toxin. Toxicology 1994; 87: 29–41.PubMedCrossRefGoogle Scholar
  75. 75.
    Koehler TM, Collier RJ. Anthrax toxin protective antigen: low pH-induced hydrophobicity and channel formation in liposomes. Mol Microbiol 1991; 5: 1501–06.PubMedCrossRefGoogle Scholar
  76. 76.
    Milne JC, Collier RJ. pH-dependent permeabilization of the plasma membrane of mammalian cells by anthrax protective antigen. Mol Microbiol 1993; 10: 647–53.PubMedCrossRefGoogle Scholar
  77. 77.
    Finkelstein A. Channels formed in phospholipid bilayer membranes by diphtheria, tetanus, botulinum, and anthrax toxin. J Physiol (Paris) 1990; 84: 188–90.Google Scholar
  78. 78.
    Zhao J, Milne JC, Collier RJ. Effect of anthrax toxin’s LF component on ion channels formed by the PA component. J Biol Chem 1995; in press.Google Scholar
  79. 79.
    Blaustein RO, Finkelstein A. Diffusion limitation in the block by symmetric tetraalkylammonium ions of anthrax toxin channels in planar phospholipid bilayer membranes. J Gen Physiol 1990; 96: 943–57.PubMedCrossRefGoogle Scholar
  80. 80.
    Blaustein RO, Finkelstein A. Voltage-dependent block of anthrax toxin channels in planar phospholipid bilayer membranes by symmetric tetraalkylammonium ions. J Gen Physiol 1990; 96: 905–19.PubMedCrossRefGoogle Scholar
  81. 81.
    Blaustein RO, Finkelstein A. Voltage-dependent block of anthrax toxin channels in planar phospholipid bilayer membranes by symmetric tetraalkylammonium ions. Effects on macroscopic conductance. J Gen Physiol 1990; 96: 921–42.PubMedCrossRefGoogle Scholar
  82. 82.
    Perelle S, Gibert M, Boquet P et al. Characterization of Clostridium perfringens iota-toxin genes and expression in Escherichia coli. Infect Immun 1993; 61: 5147–56.PubMedGoogle Scholar
  83. 83.
    Stiles BG, Wilkins TD. Clostridium perfringens iota toxin: synergism between two proteins. Toxicon 1986; 24: 767–73.PubMedCrossRefGoogle Scholar
  84. 84.
    Stiles BG, Wilkins TD. Purification and characterization of Clostridium perfringens iota toxin: dependence on two nonlinked proteins for biological activity. Infect Immun 1986; 54: 683–88.PubMedGoogle Scholar
  85. 85.
    Simpson LL, Stiles BG, Zepeda HH et al. Molecular basis for the pathological actions of Clostridium perfringens iota toxin. Infect Immun 1987; 55: 118–22.PubMedGoogle Scholar
  86. 86.
    Vandekerckhove J, Schering B, Barmann M et al. Clostridium perfringens iota toxin ADP-ribosylates skeletal muscle actin in Arg-177. FEBS Lett 1987; 225: 48–52.PubMedCrossRefGoogle Scholar
  87. 87.
    Popoff MR, Milward FW, Bancillon B et al. Purification of the Clostridium spiroforme binary toxin and activity on HEp-2 cells. Infect Immun 1989; 57: 2462–69.PubMedGoogle Scholar

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© Springer-Verlag Berlin Heidelberg 1996

Authors and Affiliations

  • Carlo Petosa
  • Robert C. Liddington

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