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Enhancement of Diphtheria Toxin Potency by Replacement of the Receptor Binding Domain with Tetanus Toxin C‐Fragment
Author(s) -
Francis Jonathan W.,
Brown Robert H.,
Figueiredo Dayse,
Remington Mary P.,
Castillo Orlando,
Schwarzschild Michael A.,
Fishman Paul S.,
Murphy John R.,
VanderSpek Johanna C.
Publication year - 2000
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.2000.0742528.x
Subject(s) - diphtheria toxin , toxin , corynebacterium diphtheriae , anthrax toxin , fusion protein , recombinant dna , cytotoxic t cell , microbiology and biotechnology , immunotoxin , chemistry , receptor , biology , diphtheria , cytotoxicity , in vitro , biochemistry , virology , vaccination , gene
This study describes the expression, purification, and characterization of a recombinant fusion toxin, DAB 389 TTC, composed of the catalytic and membrane translocation domains of diphtheria toxin (DAB 389 ) linked to the receptor binding fragment of tetanus toxin (C‐fragment). As determined by its ability to inhibit cellular protein synthesis in primary neuron cultures, DAB 389 TTC was ∼ 1,000‐fold more cytotoxic than native diphtheria toxin or the previously described fusion toxin, DAB 389 MSH. The cytotoxic effect of DAB 389 TTC on cultured cells was specific toward neuronal‐type cells and was blocked by coincubation of the chimeric toxin with tetanus antitoxin. The toxicity of DAB 389 TTC, like that of diphtheria toxin, was dependent on passage through an acidic compartment and ADP‐ribosyltransferase activity of the DAB 389 catalytic fragment. These results suggest that a catalytically inactive form of DAB 389 TTC may be useful as a nonviral vehicle to deliver exogenous proteins to the cytosolic compartment of neurons.