Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue
Abstract
Clostridium difficile is the leading cause of hospital-acquired diarrhea and pseudomembranous colitis worldwide. The organism produces two homologous toxins, TcdA and TcdB, which enter and disrupt host cell function by glucosylating and thereby inactivating key signalling molecules within the host. As a toxin-mediated disease, there has been a significant interest in identifying small molecule inhibitors of the toxins’ glucosyltransferase activities. This study was initiated as part of an effort to identify the mode of inhibition for a small molecule inhibitor of glucosyltransferase activity called apigenin. In the course of trying to get co-crystals with this inhibitor, we determined five different structures of the TcdA and TcdB glucosyltransferase domains and made use of a non-hydrolyzable UDP-glucose substrate. While we were able to visualize apigenin bound in one of our structures, the site was a crystal packing interface and not likely to explain the mode of inhibition. Nevertheless, the structure allowed us to capture an apo-state (one without the sugar nucleotide substrate) of the TcdB glycosyltransferase domain that had not been previously observed. Additionally, comparison of this structure with structures obtained in the presence of a non-hydrolyzable UDP-glucose analogue have allowed us to document multiple conformations of a C-terminal loop important formore »
- Authors:
-
- Vanderbilt Univ., Nashville, TN (United States)
- Vanderbilt Univ., Nashville, TN (United States); Vanderbilt Univ. Medical Center, Nashville, TN (United States); The Veterans Affairs Tennessee Valley Healthcare System, Nashville, TN (United States)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- National Institutes of Health (NIH); US Department of Veterans Affairs (VA)
- OSTI Identifier:
- 1438879
- Grant/Contract Number:
- AI095755; T32 GM008320; BX002943
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Structural Biology
- Additional Journal Information:
- Journal Volume: 198; Journal Issue: 3; Journal ID: ISSN 1047-8477
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Glucosyltransferase; Bacterial toxin; UDP-glucose; Clostridium difficile; Crystallography
Citation Formats
Alvin, Joseph W., and Lacy, D. Borden. Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue. United States: N. p., 2017.
Web. doi:10.1016/j.jsb.2017.04.006.
Alvin, Joseph W., & Lacy, D. Borden. Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue. United States. https://doi.org/10.1016/j.jsb.2017.04.006
Alvin, Joseph W., and Lacy, D. Borden. Wed .
"Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue". United States. https://doi.org/10.1016/j.jsb.2017.04.006. https://www.osti.gov/servlets/purl/1438879.
@article{osti_1438879,
title = {Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue},
author = {Alvin, Joseph W. and Lacy, D. Borden},
abstractNote = {Clostridium difficile is the leading cause of hospital-acquired diarrhea and pseudomembranous colitis worldwide. The organism produces two homologous toxins, TcdA and TcdB, which enter and disrupt host cell function by glucosylating and thereby inactivating key signalling molecules within the host. As a toxin-mediated disease, there has been a significant interest in identifying small molecule inhibitors of the toxins’ glucosyltransferase activities. This study was initiated as part of an effort to identify the mode of inhibition for a small molecule inhibitor of glucosyltransferase activity called apigenin. In the course of trying to get co-crystals with this inhibitor, we determined five different structures of the TcdA and TcdB glucosyltransferase domains and made use of a non-hydrolyzable UDP-glucose substrate. While we were able to visualize apigenin bound in one of our structures, the site was a crystal packing interface and not likely to explain the mode of inhibition. Nevertheless, the structure allowed us to capture an apo-state (one without the sugar nucleotide substrate) of the TcdB glycosyltransferase domain that had not been previously observed. Additionally, comparison of this structure with structures obtained in the presence of a non-hydrolyzable UDP-glucose analogue have allowed us to document multiple conformations of a C-terminal loop important for catalysis. We present our analysis of these five new structures with the hope that it will advance inhibitor design efforts for this important class of biological toxins.},
doi = {10.1016/j.jsb.2017.04.006},
journal = {Journal of Structural Biology},
number = 3,
volume = 198,
place = {United States},
year = {Wed Apr 19 00:00:00 EDT 2017},
month = {Wed Apr 19 00:00:00 EDT 2017}
}
Web of Science
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