Extension of juxtamembrane domain of diphtheria toxin receptor arrests translocation of diphtheria toxin fragment A into cytosol

Diphtheria toxin (DT) binds to the EGF-like domain of the DT receptor (DTR) , followed by internalization and translocation of the enzymatically active fragment A into the cytosol. The juxtamembrane domain (JM) of the DTR is t he linker domain connecting the transmembrane and EGF-like domains. We cons tructed mutants of DTRs with altered JMs and studied their abilities for DT intoxication. Although DTR mutants with extended JMs showed normal DT bind ing activity, the cells expressing the mutants showed both reduced transloc ation of DT fragment A into the cytosol and reduced sensitivity to DT, when compared with cells expressing wild-type DTR. These results indicate that the JM contributes to DT intoxication by providing a space appropriate for the interaction of DT with the cell membrane. The present study also indica tes that consideration of epitopes of an immunotoxins would be an important factor in the design of potent immunotoxins. (C) 2001 Academic Press.