Proteolytic activation of bacterial toxins by eukaryotic cells is performed by furin and by additional cellular proteases - PubMed (original) (raw)

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

V M Gordon et al. Infect Immun. 1995 Jan.

Abstract

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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References

    1. FEBS Lett. 1993 Dec 20;336(1):65-9 - PubMed
    1. J Biol Chem. 1991 Sep 15;266(26):17376-81 - PubMed
    1. Infect Immun. 1994 Feb;62(2):333-40 - PubMed
    1. J Virol. 1994 Jun;68(6):4075-9 - PubMed
    1. J Cell Sci. 1994 Mar;107 ( Pt 3):719-26 - PubMed

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