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Infect. Immun., 01 1995, 82-87, Vol 63, No. 1
Copyright © 1995, American Society for Microbiology

Proteolytic activation of bacterial toxins by eukaryotic cells is performed by furin and by additional cellular proteases

VM Gordon, KR Klimpel, N Arora, MA Henderson and SH Leppla
Laboratory of Microbial Ecology, National Institute of Dental Research, Bethesda, Maryland 20892.

Before intoxication can occur, anthrax toxin protective antigen (PA), Pseudomonas exotoxin A (PE), and diphtheria toxin (DT) must be activated by proteolytic cleavage at specific amino acid sequences. Previously, it was shown that PA and DT can be activated by furin. In Chinese hamster ovary (CHO) cells, wild-type (RKKR) and cleavage site mutants of PA, each administered with a modified form of anthrax toxin lethal factor (the N terminus of lethal factor fused to PE domain III), had the following potencies: RKKR (wild type) (concentration causing 50% cell death [EC50] = 12 ng/ml) > or = RAAR (EC50 = 18 ng/ml) > FTKR (EC50 = 24 ng/ml) > STRR (EC50 = 49 ng/ml). In vitro cleavage of PA and cleavage site mutants of PA by furin demonstrated that native PA (RKKR) and PA with the cleavage sequence RAAR are substrates for furin. To characterize eukaryotic proteases that play a role in activating bacterial toxins, furin-deficient CHO cells were selected after chemical mutagenesis. Furin-deficient cells were resistant to PE, whose cleavage site, RQPR, constitutes a furin recognition site and to all PA cleavage site mutants, but were sensitive to DT (EC50 = 2.9 ng/ml) and PA (EC50 = 23 ng/ml), whose respective cleavage sites, RKKR and RVRR, contain additional basic residues. Furin-deficient cells that were transfected with the furin gene regained sensitivity to PE and PA cleavage site mutants. These studies provide evidence that furin can activate the three toxins and that one or more additional proteases contribute to the activation of DT and PA.


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  • Farchaus, J. W., Ribot, W. J., Jendrek, S., Little, S. F. (1998). Fermentation, Purification, and Characterization of Protective Antigen from a Recombinant, Avirulent Strain of Bacillus anthracis. Appl. Environ. Microbiol. 64: 982-991 [Abstract] [Full Text]  
  • Michael Lord, J., Roberts, L. M. (1998). Toxin Entry: Retrograde Transport through the Secretory Pathway. JCB 140: 733-736 [Full Text]  
  • Ballard, J. D., Doling, A. M., Beauregard, K., Collier, R. J., Starnbach, M. N. (1998). Anthrax Toxin-Mediated Delivery In Vivo and In Vitro of a Cytotoxic T-Lymphocyte Epitope from Ovalbumin. Infect. Immun. 66: 615-619 [Abstract] [Full Text]  
  • Liu, G., Thomas, L., Warren, R. A., Enns, C. A., Cunningham, C. C., Hartwig, J. H., Thomas, G. (1997). Cytoskeletal Protein ABP-280 Directs the Intracellular Trafficking of Furin and Modulates Proprotein Processing in the Endocytic Pathway. JCB 139: 1719-1733 [Abstract] [Full Text]  
  • Frankel, A. E., Hall, P. D., Burbage, C., Vesely, J., Willingham, M., Bhalla, K., Kreitman, R. J. (1997). Modulation of the Apoptotic Response of Human Myeloid Leukemia Cells to a Diphtheria Toxin Granulocyte-Macrophage Colony-Stimulating Factor Fusion Protein. Blood 90: 3654-3661 [Abstract] [Full Text]  


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