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 »
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- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1581008
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- AC02-76SF00515
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- 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
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- Proceedings of the National Academy of Sciences
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- United States
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- 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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