Structure of the cell-binding component of the Clostridium difficile binary toxin reveals a di-heptamer macromolecular assembly
Abstract
TargetingClostridium difficileinfection is challenging because treatment options are limited, and high recurrence rates are common. One reason for this is that hypervirulentC. difficilestrains often have a binary toxin termed theC. difficiletoxin, in addition to the enterotoxins TsdA and TsdB. TheC. difficiletoxin 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 (SymCDTb; 3.14 Å) and an asymmetric form (AsymCDTb; 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). ForAsymCDTb, a Ca2+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 targetmore »
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- Publication Date:
- Research Org.:
- SLAC National Accelerator Lab., Menlo Park, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1581008
- Alternate Identifier(s):
- OSTI ID: 1625064
- 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
- Subject:
- Science & Technology - Other Topics
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 = {TargetingClostridium difficileinfection is challenging because treatment options are limited, and high recurrence rates are common. One reason for this is that hypervirulentC. difficilestrains often have a binary toxin termed theC. difficiletoxin, in addition to the enterotoxins TsdA and TsdB. TheC. difficiletoxin 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 (SymCDTb; 3.14 Å) and an asymmetric form (AsymCDTb; 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). ForAsymCDTb, a Ca2+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 ofC. 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}
}
DOI: 10.1073/pnas.1919490117
Web of Science
Figures / Tables:

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