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    Masking autoprocessing of Clostridium difficile toxin A by the C-terminus combined repetitive oligo peptides.
    • Authors:
    • Yongrong Zhang
      Tufts Cummings School of Veterinary Medicine
      United States
      Therwa Hamza
      University of Maryland Dental School
      Baltimore | United States
      Si Gao
      School of Pharmaceutical Sciences
      China
      Hanping Feng
      Tufts University Cummings School of Veterinary Medicine
      United States
    Biochem Biophys Res Commun 2015 Apr 26;459(2):259-63. Epub 2015 Feb 26.
    Department of Microbial Pathogenesis, University of Maryland Dental School, Baltimore, MD 21201, USA. Electronic address:
    Clostridium difficile toxin A and B (TcdA and TcdB) are the major virulence factors of the bacterium, both of which consist of two enzymatic domains: an effector glucosyltransferase domain (GTD) and a cysteine protease domain (CPD) responsible for autocleavage and release of GTD. Although the CPDs from both toxins share a similar structure and mechanism of hexakisphosphate (InsP6)-induced activation, TcdA is substantially less sensitive to the autocleavage as compared with TcdB. In this study, we provided evidence of inter-domain regulation of CPD activity of TcdA and its autoprocessing. The C-terminus combined repetitive oligo peptides (CROPs) of TcdA reduced the accessibility of TcdB CPD to its substrate in a chimeric toxin TxB-Ar, consequently blocking autoprocessing. Moreover, interference of antibodies with the CROPs of full-length TcdA efficiently enhanced its GTD release. In conclusion, by utilizing chimeric toxins and specific antibodies, we identified that the CROPs of TcdA plays a crucial role in controlling the InsP6-mediated activation of CPD and autocleavage of GTD. Our data provides insights on the molecular mode of action of the C. difficile toxins.

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