Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue

Alvin, Joseph W.; Lacy, D. Borden

doi:10.1016/j.jsb.2017.04.006

Title: Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue

Journal Article · Wed Apr 19 00:00:00 EDT 2017 · Journal of Structural Biology

DOI:https://doi.org/10.1016/j.jsb.2017.04.006· OSTI ID:1438879

Alvin, Joseph W. ^[1];

^[2]

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)

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.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); US Department of Veterans Affairs (VA)

Grant/Contract Number:: AI095755; T32 GM008320; BX002943

OSTI ID:: 1438879

Journal Information:: Journal of Structural Biology, Vol. 198, Issue 3; ISSN 1047-8477

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

Citation Metrics:

Cited by: 10 works

Citation information provided by
Web of Science

References (26)

PHENIX: a comprehensive Python-based system for macromolecular structure solution Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221 https://doi.org/10.1107/S0907444909052925	journal	January 2010
Structures and mechanisms of glycosyltransferases Breton, Christelle; Šnajdrová, Lenka; Jeanneau, Charlotte Glycobiology, Vol. 16, Issue 2 https://doi.org/10.1093/glycob/cwj016	journal	July 2005
A Common Motif of Eukaryotic Glycosyltransferases Is Essential for the Enzyme Activity of Large Clostridial Cytotoxins Busch, Christian; Hofmann, Fred; Selzer, Jörg Journal of Biological Chemistry, Vol. 273, Issue 31 https://doi.org/10.1074/jbc.273.31.19566	journal	July 1998
Involvement of a Conserved Tryptophan Residue in the UDP-Glucose Binding of Large Clostridial Cytotoxin Glycosyltransferases Busch, Christian; Hofmann, Fred; Gerhard, Ralf Journal of Biological Chemistry, Vol. 275, Issue 18 https://doi.org/10.1074/jbc.275.18.13228	journal	May 2000
Toxins A and B from Clostridium difficile differ with respect to enzymatic potencies, cellular substrate specificities, and surface binding to cultured cells. Chaves-Olarte, E.; Weidmann, M.; Eichel-Streiber, C. Journal of Clinical Investigation, Vol. 100, Issue 7 https://doi.org/10.1172/JCI119698	journal	October 1997
Crystal structure of Clostridium difficile toxin A Chumbler, Nicole M.; Rutherford, Stacey A.; Zhang, Zhifen Nature Microbiology, Vol. 1, Issue 1 https://doi.org/10.1038/nmicrobiol.2015.2	journal	January 2016
Clostridium difficile Toxins A and B Are Cation-dependent UDP-glucose Hydrolases with Differing Catalytic Activities Ciesla, William P.; Bobak, David A. Journal of Biological Chemistry, Vol. 273, Issue 26 https://doi.org/10.1074/jbc.273.26.16021	journal	June 1998
The structure of Clostridium difficile toxin A glucosyltransferase domain bound to Mn ²⁺ and UDP provides insights into glucosyltransferase activity and product release: Structure of TcdA-GT bound to Mn ²⁺ and UDP D’Urzo, Nunzia; Malito, Enrico; Biancucci, Marco FEBS Journal, Vol. 279, Issue 17 https://doi.org/10.1111/j.1742-4658.2012.08688.x	journal	July 2012
Features and development of Coot Emsley, P.; Lohkamp, B.; Scott, W. G. Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4 https://doi.org/10.1107/S0907444910007493	journal	March 2010
The Donor Subsite of Trehalose-6-phosphate Synthase: BINARY COMPLEXES WITH UDP-GLUCOSE AND UDP-2-DEOXY-2-FLUORO-GLUCOSE AT 2 Å RESOLUTION Gibson, Robert P.; Tarling, Chris A.; Roberts, Shirley Journal of Biological Chemistry, Vol. 279, Issue 3 https://doi.org/10.1074/jbc.M307643200	journal	October 2003
Crystal structure of UDP-glucose:anthocyanidin 3- O -glucosyltransferase from Clitoria ternatea Hiromoto, Takeshi; Honjo, Eijiro; Tamada, Taro Journal of Synchrotron Radiation, Vol. 20, Issue 6 https://doi.org/10.1107/S0909049513020712	journal	September 2013
Sequencing and analysis of the gene encoding the α-toxin of Clostridium novyi proves its homology to toxins A and B of Clostridium difficile Hofmann, Fred; Herrmann, Andrea; Habermann, Ernst Molecular and General Genetics MGG, Vol. 247, Issue 6 https://doi.org/10.1007/BF00290398	journal	November 1995
Structure and mode of action of clostridial glucosylating toxins: the ABCD model Jank, Thomas; Aktories, Klaus Trends in Microbiology, Vol. 16, Issue 5 https://doi.org/10.1016/j.tim.2008.01.011	journal	May 2008
Clostridium difficile Glucosyltransferase Toxin B-essential Amino Acids for Substrate Binding Jank, Thomas; Giesemann, Torsten; Aktories, Klaus Journal of Biological Chemistry, Vol. 282, Issue 48 https://doi.org/10.1074/jbc.M703138200	journal	September 2007
Glucosylation of Rho proteins by Clostridium difficile toxin B Just, I.; Selzer, J.; Wilm, M. Nature, Vol. 375, Issue 6531 https://doi.org/10.1038/375500a0	journal	June 1995
XDS Kabsch, Wolfgang Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2 https://doi.org/10.1107/S0907444909047337	journal	January 2010
Glycosylating Toxin of Clostridium perfringens Nagahama, Masahiro; Oda, Masataka; Kobayashi, Keiko Glycosylation https://doi.org/10.5772/48112	book	September 2012
[20] Processing of X-ray diffraction data collected in oscillation mode Otwinowski, Zbyszek; Minor, Wladek Macromolecular Crystallography Part A, 307-326 https://doi.org/10.1016/S0076-6879(97)76066-X	book	January 1997
Structural Determinants of Clostridium difficile Toxin A Glucosyltransferase Activity Pruitt, Rory N.; Chumbler, Nicole M.; Rutherford, Stacey A. Journal of Biological Chemistry, Vol. 287, Issue 11 https://doi.org/10.1074/jbc.M111.298414	journal	January 2012
Autocatalytic cleavage of Clostridium difficile toxin B Reineke, Jessica; Tenzer, Stefan; Rupnik, Maja Nature, Vol. 446, Issue 7134 https://doi.org/10.1038/nature05622	journal	March 2007
Structural Basis for the Function of Clostridium difficile Toxin B Reinert, Dirk J.; Jank, Thomas; Aktories, Klaus Journal of Molecular Biology, Vol. 351, Issue 5 https://doi.org/10.1016/j.jmb.2005.06.071	journal	September 2005
Mutational Analysis of the Enzymatic Domain of Clostridium difficile Toxin B Reveals Novel Inhibitors of the Wild-Type Toxin Spyres, Lea M.; Daniel, Jeremy; Hensley, Amy Infection and Immunity, Vol. 71, Issue 6 https://doi.org/10.1128/IAI.71.6.3294-3301.2003	journal	June 2003
Small Molecule Inhibitors of Clostridium difficile Toxin B-Induced Cellular Damage Tam, John; Beilhartz, Greg L.; Auger, Anick Chemistry & Biology, Vol. 22, Issue 2 https://doi.org/10.1016/j.chembiol.2014.12.010	journal	February 2015
Application of Mutated Clostridium difficile Toxin A for Determination of Glucosyltransferase-Dependent Effects Teichert, M.; Tatge, H.; Schoentaube, J. Infection and Immunity, Vol. 74, Issue 10 https://doi.org/10.1128/IAI.00545-06	journal	September 2006
Chimeric Glycosyltransferases for the Generation of Hybrid Glycopeptides Truman, Andrew W.; Dias, Marcio V. B.; Wu, Shu Chemistry & Biology, Vol. 16, Issue 6 https://doi.org/10.1016/j.chembiol.2009.04.013	journal	June 2009
Conformational Changes and Reaction of Clostridial Glycosylating Toxins Ziegler, Mathias O. P.; Jank, Thomas; Aktories, Klaus Journal of Molecular Biology, Vol. 377, Issue 5 https://doi.org/10.1016/j.jmb.2007.12.065	journal	April 2008

Cited By (1)

The role of toxins in Clostridium difficile infection Chandrasekaran, Ramyavardhanee; Lacy, D. Borden FEMS Microbiology Reviews, Vol. 41, Issue 6 https://doi.org/10.1093/femsre/fux048	journal	October 2017

Similar Records

Inhibition of Clostridium difficile TcdA and TcdB toxins with transition state analogues

Journal Article · Mon Nov 01 00:00:00 EDT 2021 · Nature Communications · OSTI ID:1438879

Paparella, Ashleigh S.; Aboulache, Briana L.; Harijan, Rajesh K.; +3 more

Structural Determinants of Clostridium difficile Toxin A Glucosyltransferase Activity

Journal Article · Wed Mar 28 00:00:00 EDT 2012 · Journal of Biological Chemistry · OSTI ID:1438879

Pruitt, Rory N; Chumbler, Nicole M; Rutherford, Stacey A; +4 more

Crystal structure of Clostridium difficile toxin A

Journal Article · Mon Jan 11 00:00:00 EST 2016 · Nature Microbiology · OSTI ID:1438879

Chumbler, Nicole M.; Rutherford, Stacey A.; Zhang, Zhifen; +7 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Glucosyltransferase
Bacterial toxin
UDP-glucose
Clostridium difficile
Crystallography

Title: Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue

Citation Formats

References (26)

Cited By (1)

Similar Records

Related Subjects