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Title: Structure of the cell-binding component of the Clostridium difficile binary toxin reveals a di-heptamer macromolecular assembly

Abstract

Targeting Clostridium difficile infection is challenging because treatment options are limited, and high recurrence rates are common. One reason for this is that hypervirulent C. difficile strains often have a binary toxin termed the C. difficile toxin, in addition to the enterotoxins TsdA and TsdB. The C. difficile toxin has an enzymatic component, termed CDTa, and a pore-forming or delivery subunit termed CDTb. CDTb was characterized here using a combination of single-particle cryoelectron microscopy, X-ray crystallography, NMR, and other biophysical methods. In the absence of CDTa, 2 di-heptamer structures for activated CDTb (1.0 MDa) were solved at atomic resolution, including a symmetric ( Sym CDTb; 3.14 Å) and an asymmetric form ( Asym CDTb; 2.84 Å). Roles played by 2 receptor-binding domains of activated CDTb were of particular interest since the receptor-binding domain 1 lacks sequence homology to any other known toxin, and the receptor-binding domain 2 is completely absent in other well-studied heptameric toxins (i.e., anthrax). For Asym CDTb, a Ca 2+ binding site was discovered in the first receptor-binding domain that is important for its stability, and the second receptor-binding domain was found to be critical for host cell toxicity and the di-heptamer fold for both forms ofmore » activated CDTb. Together, these studies represent a starting point for developing structure-based drug-design strategies to target the most severe strains of C. difficile .« less

Authors:
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Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1581008
Grant/Contract Number:  
AC02-76SF00515
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 117 Journal Issue: 2; Journal ID: ISSN 0027-8424
Publisher:
Proceedings of the National Academy of Sciences
Country of Publication:
United States
Language:
English

Citation Formats

Xu, Xingjian, Godoy-Ruiz, Raquel, Adipietro, Kaylin A., Peralta, Christopher, Ben-Hail, Danya, Varney, Kristen M., Cook, Mary E., Roth, Braden M., Wilder, Paul T., Cleveland, Thomas, Grishaev, Alexander, Neu, Heather M., Michel, Sarah L. J., Yu, Wenbo, Beckett, Dorothy, Rustandi, Richard R., Lancaster, Catherine, Loughney, John W., Kristopeit, Adam, Christanti, Sianny, Olson, Jessica W., MacKerell, Alexander D., Georges, Amedee des, Pozharski, Edwin, and Weber, David J. Structure of the cell-binding component of the Clostridium difficile binary toxin reveals a di-heptamer macromolecular assembly. United States: N. p., 2020. Web. doi:10.1073/pnas.1919490117.
Xu, Xingjian, Godoy-Ruiz, Raquel, Adipietro, Kaylin A., Peralta, Christopher, Ben-Hail, Danya, Varney, Kristen M., Cook, Mary E., Roth, Braden M., Wilder, Paul T., Cleveland, Thomas, Grishaev, Alexander, Neu, Heather M., Michel, Sarah L. J., Yu, Wenbo, Beckett, Dorothy, Rustandi, Richard R., Lancaster, Catherine, Loughney, John W., Kristopeit, Adam, Christanti, Sianny, Olson, Jessica W., MacKerell, Alexander D., Georges, Amedee des, Pozharski, Edwin, & Weber, David J. Structure of the cell-binding component of the Clostridium difficile binary toxin reveals a di-heptamer macromolecular assembly. United States. doi:10.1073/pnas.1919490117.
Xu, Xingjian, Godoy-Ruiz, Raquel, Adipietro, Kaylin A., Peralta, Christopher, Ben-Hail, Danya, Varney, Kristen M., Cook, Mary E., Roth, Braden M., Wilder, Paul T., Cleveland, Thomas, Grishaev, Alexander, Neu, Heather M., Michel, Sarah L. J., Yu, Wenbo, Beckett, Dorothy, Rustandi, Richard R., Lancaster, Catherine, Loughney, John W., Kristopeit, Adam, Christanti, Sianny, Olson, Jessica W., MacKerell, Alexander D., Georges, Amedee des, Pozharski, Edwin, and Weber, David J. Thu . "Structure of the cell-binding component of the Clostridium difficile binary toxin reveals a di-heptamer macromolecular assembly". United States. doi:10.1073/pnas.1919490117.
@article{osti_1581008,
title = {Structure of the cell-binding component of the Clostridium difficile binary toxin reveals a di-heptamer macromolecular assembly},
author = {Xu, Xingjian and Godoy-Ruiz, Raquel and Adipietro, Kaylin A. and Peralta, Christopher and Ben-Hail, Danya and Varney, Kristen M. and Cook, Mary E. and Roth, Braden M. and Wilder, Paul T. and Cleveland, Thomas and Grishaev, Alexander and Neu, Heather M. and Michel, Sarah L. J. and Yu, Wenbo and Beckett, Dorothy and Rustandi, Richard R. and Lancaster, Catherine and Loughney, John W. and Kristopeit, Adam and Christanti, Sianny and Olson, Jessica W. and MacKerell, Alexander D. and Georges, Amedee des and Pozharski, Edwin and Weber, David J.},
abstractNote = {Targeting Clostridium difficile infection is challenging because treatment options are limited, and high recurrence rates are common. One reason for this is that hypervirulent C. difficile strains often have a binary toxin termed the C. difficile toxin, in addition to the enterotoxins TsdA and TsdB. The C. difficile toxin has an enzymatic component, termed CDTa, and a pore-forming or delivery subunit termed CDTb. CDTb was characterized here using a combination of single-particle cryoelectron microscopy, X-ray crystallography, NMR, and other biophysical methods. In the absence of CDTa, 2 di-heptamer structures for activated CDTb (1.0 MDa) were solved at atomic resolution, including a symmetric ( Sym CDTb; 3.14 Å) and an asymmetric form ( Asym CDTb; 2.84 Å). Roles played by 2 receptor-binding domains of activated CDTb were of particular interest since the receptor-binding domain 1 lacks sequence homology to any other known toxin, and the receptor-binding domain 2 is completely absent in other well-studied heptameric toxins (i.e., anthrax). For Asym CDTb, a Ca 2+ binding site was discovered in the first receptor-binding domain that is important for its stability, and the second receptor-binding domain was found to be critical for host cell toxicity and the di-heptamer fold for both forms of activated CDTb. Together, these studies represent a starting point for developing structure-based drug-design strategies to target the most severe strains of C. difficile .},
doi = {10.1073/pnas.1919490117},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 2,
volume = 117,
place = {United States},
year = {2020},
month = {1}
}

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DOI: 10.1073/pnas.1919490117

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