Abstract
Besides two large cytotoxins (TcdA and TcdB), certain Clostridioides difficile strains also produce a binary toxin, called C. difficile toxin (CDT) composed of an enzymatic subunit involved in actin ADP-ribosylation (CDTa) and translocation pore (CDTb) that delivers CDTa into host cells through receptor-mediated endocytosis. CDTb is proposed to be a di-heptamer, but its physiological heptameric structure has not been reported to date. Here, we report the CDTa-bound CDTb-pore (heptamer) as a physiological complexes using cryo-EM. The high-resolution structure of the CDTa-bound CDTb-pore at 2.56-Å resolution revealed that CDTa binding to CDTb-pore induces partial unfolding and tilting of the first CDTa a-helix, and the translocation. In the CDTb-pore, the NSS-loop exists in “in” and “out” conformations, suggesting their involvement in substrate translocation through formation of weak, non-specific interactions. This structural information provides insights into drug design against hypervirulent C. difficile strains.
Cryo-EM structures of the translocational binary toxin complex CDTa-bound CDTb-pore from Clostridioides difficile
