Inhibition of Clostridium difficile TcdA and TcdB toxins with transition state analogues
Abstract
Clostridium difficile causes life-threatening diarrhea and is the leading cause of healthcare-associated bacterial infections in the United States. TcdA and TcdB bacterial toxins are primary determinants of disease pathogenesis and are attractive therapeutic targets. TcdA and TcdB contain domains that use UDP-glucose to glucosylate and inactivate host Rho GTPases, resulting in cytoskeletal changes causing cell rounding and loss of intestinal integrity. Transition state analysis revealed glucocationic character for the TcdA and TcdB transition states. We identified transition state analogue inhibitors and characterized them by kinetic, thermodynamic and structural analysis. Iminosugars, isofagomine and noeuromycin mimic the transition state and inhibit both TcdA and TcdB by forming ternary complexes with Tcd and UDP, a product of the TcdA- and TcdB-catalyzed reactions. Both iminosugars prevent TcdA- and TcdB-induced cytotoxicity in cultured mammalian cells by preventing glucosylation of Rho GTPases. Iminosugar transition state analogues of the Tcd toxins show potential as therapeutics for C. difficile pathology.
- Authors:
-
[1];
[2];
[1];
[3];
[1]
- Albert Einstein College of Medicine, Bronx, NY (United States)
- Albert Einstein College of Medicine, Bronx, NY (United States); Univ. of Colorado, Boulder, CO (United States)
- Victoria Univ. of Wellington, Lower Hutt (New Zealand)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS); Brookhaven National Lab. (BNL), Upton, NY (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Institutes of Health, National Institute of General Medical Sciences (NIGMS); USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1829876
- Alternate Identifier(s):
- OSTI ID: 1875645
- Report Number(s):
- BNL-223143-2022-JACI
Journal ID: ISSN 2041-1723; APS_263792
- Grant/Contract Number:
- AC02-06CH11357; SC0012704; P30GM133893; GM041916; AI150971; S10 OD020068
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Nature Communications
- Additional Journal Information:
- Journal Volume: 12; Journal Issue: 1; Journal ID: ISSN 2041-1723
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 99 GENERAL AND MISCELLANEOUS; Enzymes; Transferases; X-ray crystallography
Citation Formats
Paparella, Ashleigh S., Aboulache, Briana L., Harijan, Rajesh K., Potts, Kathryn S., Tyler, Peter C., and Schramm, Vern L. Inhibition of Clostridium difficile TcdA and TcdB toxins with transition state analogues. United States: N. p., 2021.
Web. doi:10.1038/s41467-021-26580-6.
Paparella, Ashleigh S., Aboulache, Briana L., Harijan, Rajesh K., Potts, Kathryn S., Tyler, Peter C., & Schramm, Vern L. Inhibition of Clostridium difficile TcdA and TcdB toxins with transition state analogues. United States. https://doi.org/10.1038/s41467-021-26580-6
Paparella, Ashleigh S., Aboulache, Briana L., Harijan, Rajesh K., Potts, Kathryn S., Tyler, Peter C., and Schramm, Vern L. Mon .
"Inhibition of Clostridium difficile TcdA and TcdB toxins with transition state analogues". United States. https://doi.org/10.1038/s41467-021-26580-6. https://www.osti.gov/servlets/purl/1829876.
@article{osti_1829876,
title = {Inhibition of Clostridium difficile TcdA and TcdB toxins with transition state analogues},
author = {Paparella, Ashleigh S. and Aboulache, Briana L. and Harijan, Rajesh K. and Potts, Kathryn S. and Tyler, Peter C. and Schramm, Vern L.},
abstractNote = {Clostridium difficile causes life-threatening diarrhea and is the leading cause of healthcare-associated bacterial infections in the United States. TcdA and TcdB bacterial toxins are primary determinants of disease pathogenesis and are attractive therapeutic targets. TcdA and TcdB contain domains that use UDP-glucose to glucosylate and inactivate host Rho GTPases, resulting in cytoskeletal changes causing cell rounding and loss of intestinal integrity. Transition state analysis revealed glucocationic character for the TcdA and TcdB transition states. We identified transition state analogue inhibitors and characterized them by kinetic, thermodynamic and structural analysis. Iminosugars, isofagomine and noeuromycin mimic the transition state and inhibit both TcdA and TcdB by forming ternary complexes with Tcd and UDP, a product of the TcdA- and TcdB-catalyzed reactions. Both iminosugars prevent TcdA- and TcdB-induced cytotoxicity in cultured mammalian cells by preventing glucosylation of Rho GTPases. Iminosugar transition state analogues of the Tcd toxins show potential as therapeutics for C. difficile pathology.},
doi = {10.1038/s41467-021-26580-6},
journal = {Nature Communications},
number = 1,
volume = 12,
place = {United States},
year = {Mon Nov 01 00:00:00 EDT 2021},
month = {Mon Nov 01 00:00:00 EDT 2021}
}
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Works referenced in this record:
Glucosylation of Rho proteins by Clostridium difficile toxin B
journal, June 1995
- Just, I.; Selzer, J.; Wilm, M.
- Nature, Vol. 375, Issue 6531
Clostridium difficile Toxin Biology
journal, September 2017
- Aktories, Klaus; Schwan, Carsten; Jank, Thomas
- Annual Review of Microbiology, Vol. 71, Issue 1
Refinement of Macromolecular Structures by the Maximum-Likelihood Method
journal, May 1997
- Murshudov, G. N.; Vagin, A. A.; Dodson, E. J.
- Acta Crystallographica Section D Biological Crystallography, Vol. 53, Issue 3
Mechanistic evidence for a front-side, SNi-type reaction in a retaining glycosyltransferase
journal, August 2011
- Lee, Seung Seo; Hong, Sung You; Errey, James C.
- Nature Chemical Biology, Vol. 7, Issue 9
Transition State Analysis Using Multiple Kinetic Isotope Effects: Mechanisms of Enzymatic and Non-Enzymatic Glycoside Hydrolysis and Transfer
journal, September 2003
- Berti, Paul J.; Tanaka, Kelly S. E.
- ChemInform, Vol. 34, Issue 35
Difference in the biological effects of Clostridium difficile toxin B in proliferating and non-proliferating cells
journal, January 2011
- Lica, Marta; Schulz, Florian; Schelle, Ilona
- Naunyn-Schmiedeberg's Archives of Pharmacology, Vol. 383, Issue 3
Clostridium difficile Glucosyltransferase Toxin B-essential Amino Acids for Substrate Binding
journal, September 2007
- Jank, Thomas; Giesemann, Torsten; Aktories, Klaus
- Journal of Biological Chemistry, Vol. 282, Issue 48
Bacterial protein toxins that modify host regulatory GTPases
journal, June 2011
- Aktories, Klaus
- Nature Reviews Microbiology, Vol. 9, Issue 7
Kinetics of oxygen exchange at the anomeric carbon atom of D-glucose and D-erythrose using the oxygen-18 isotope effect in carbon-13 nuclear magnetic resonance spectroscopy
journal, December 1982
- Risley, John M.; Van Etten, Robert L.
- Biochemistry, Vol. 21, Issue 25
Toward a Detailed Understanding of Base Excision Repair Enzymes: Transition State and Mechanistic Analyses of N -Glycoside Hydrolysis and N- Glycoside Transfer
journal, February 2006
- Berti, Paul J.; McCann, Joe A. B.
- Chemical Reviews, Vol. 106, Issue 2
A small-molecule antivirulence agent for treating Clostridium difficile infection
journal, September 2015
- Bender, Kristina Oresic; Garland, Megan; Ferreyra, Jessica A.
- Science Translational Medicine, Vol. 7, Issue 306
Update on Antimicrobial Resistance in Clostridium difficile: Resistance Mechanisms and Antimicrobial Susceptibility Testing
journal, July 2017
- Peng, Zhong; Jin, Dazhi; Kim, Hyeun Bum
- Journal of Clinical Microbiology, Vol. 55, Issue 7
The role of toxin A and toxin B in Clostridium difficile infection
journal, September 2010
- Kuehne, Sarah A.; Cartman, Stephen T.; Heap, John T.
- Nature, Vol. 467, Issue 7316
Complete kinetic isotope effect description of transition states for acid-catalyzed hydrolyses of methyl .alpha.- and .beta.-glucopyranosides
journal, November 1986
- Bennet, Andrew J.; Sinnott, Michael L.
- Journal of the American Chemical Society, Vol. 108, Issue 23
Clostridium difficile toxin glucosyltransferase domains in complex with a non-hydrolyzable UDP-glucose analogue
journal, June 2017
- Alvin, Joseph W.; Lacy, D. Borden
- Journal of Structural Biology, Vol. 198, Issue 3
Structural Basis for the Function of Clostridium difficile Toxin B
journal, September 2005
- Reinert, Dirk J.; Jank, Thomas; Aktories, Klaus
- Journal of Molecular Biology, Vol. 351, Issue 5
Bisubstrate Analogues as Glycosyltransferase Inhibitors
journal, January 2009
- Izumi, Masayuki; Yuasa, Hideya; Hashimoto, Hironobu
- Current Topics in Medicinal Chemistry, Vol. 9, Issue 1
Host-targeted niclosamide inhibits C. difficile virulence and prevents disease in mice without disrupting the gut microbiota
journal, December 2018
- Tam, John; Hamza, Therwa; Ma, Bing
- Nature Communications, Vol. 9, Issue 1
Clostridium difficile infection: Toxins and non-toxin virulence factors, and their contributions to disease establishment and host response
journal, March 2012
- Vedantam, Gayatri; Clark, Andrew; Chu, Michele
- Gut Microbes, Vol. 3, Issue 2
Glycosidase inhibition: assessing mimicry of the transition state
journal, January 2010
- Gloster, Tracey M.; Davies, Gideon J.
- Org. Biomol. Chem., Vol. 8, Issue 2
Recent Developments of Transition-State Analogue Glycosidase Inhibitors of Non-Natural Product Origin
journal, February 2002
- Lillelund, Vinni H.; Jensen, Henrik H.; Liang, Xifu
- Chemical Reviews, Vol. 102, Issue 2
Burden of Clostridium difficile on the Healthcare System
journal, August 2012
- Dubberke, Erik R.; Olsen, Margaret A.
- Clinical Infectious Diseases, Vol. 55, Issue suppl_2
Clostridium difficile Toxins A and B Are Cation-dependent UDP-glucose Hydrolases with Differing Catalytic Activities
journal, June 1998
- Ciesla, William P.; Bobak, David A.
- Journal of Biological Chemistry, Vol. 273, Issue 26
Isofagomine In Vivo Effects in a Neuronopathic Gaucher Disease Mouse
journal, April 2011
- Sun, Ying; Ran, Huimin; Liou, Benjamin
- PLoS ONE, Vol. 6, Issue 4
The CCP4 suite programs for protein crystallography
journal, September 1994
- ,
- Acta Crystallographica Section D Biological Crystallography, Vol. 50, Issue 5, p. 760-763
Virulence of new variant strains of Clostridium difficile producing only toxin A or binary toxin in the hamster model
journal, November 2019
- Marvaud, J. -C.; Quevedo-Torres, S.; Eckert, C.
- New Microbes and New Infections, Vol. 32
The role of toxin A and toxin B in Clostridium difficile-associated disease: Past and present perspectives
journal, January 2010
- Carter, Glen P.; Rood, Julian I.; Lyras, Dena
- Gut Microbes, Vol. 1, Issue 1
Inhibition of the glucosyltransferase activity of clostridial Rho/Ras-glucosylating toxins by castanospermine
journal, May 2008
- Jank, Thomas; Ziegler, Mathias O. P.; Schulz, Georg E.
- FEBS Letters, Vol. 582, Issue 15
Enzymatic Transition States and Transition State Analog Design
journal, June 1998
- Schramm, Vern L.
- Annual Review of Biochemistry, Vol. 67, Issue 1
Differential Effects of Clostridium difficile Toxins on Tissue-Cultured Cells
journal, June 1982
- Donta, Sam T.; Sullivan, Nadine; Wilkins, Tracy D.
- Journal of Clinical Microbiology, Vol. 15, Issue 6
Clostridium difficile Infection
journal, April 2015
- Leffler, Daniel A.; Lamont, J. Thomas
- New England Journal of Medicine, Vol. 372, Issue 16
Synthesis and SAR studies of novel benzodiazepinedione-based inhibitors of Clostridium difficile (C. difficile) toxin B (TcdB)
journal, December 2018
- Letourneau, Jeffrey J.; Stroke, Ilana L.; Hilbert, David W.
- Bioorganic & Medicinal Chemistry Letters, Vol. 28, Issue 23-24
Toxin B is essential for virulence of Clostridium difficile
journal, March 2009
- Lyras, Dena; O’Connor, Jennifer R.; Howarth, Pauline M.
- Nature, Vol. 458, Issue 7242
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
journal, July 2012
- D’Urzo, Nunzia; Malito, Enrico; Biancucci, Marco
- FEBS Journal, Vol. 279, Issue 17
iMOSFLM : a new graphical interface for diffraction-image processing with MOSFLM
journal, March 2011
- Battye, T. Geoff G.; Kontogiannis, Luke; Johnson, Owen
- Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
Bezlotoxumab: First Global Approval
journal, November 2016
- Markham, Anthony
- Drugs, Vol. 76, Issue 18
The Expression of Isotope Effects on Enzyme-Catalyzed Reactions
journal, June 1981
- Northrop, D. B.
- Annual Review of Biochemistry, Vol. 50, Issue 1
Hyperconjugation and the angular dependence of .beta.-deuterium isotope effects
journal, June 1977
- Sunko, D. E.; Szele, Ivanka; Hehre, Warren J.
- Journal of the American Chemical Society, Vol. 99, Issue 15
MolProbity : all-atom structure validation for macromolecular crystallography
journal, December 2009
- Chen, Vincent B.; Arendall, W. Bryan; Headd, Jeffrey J.
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1
Features and development of Coot
journal, March 2010
- Emsley, P.; Lohkamp, B.; Scott, W. G.
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
Structural Determinants of Clostridium difficile Toxin A Glucosyltransferase Activity
journal, January 2012
- Pruitt, Rory N.; Chumbler, Nicole M.; Rutherford, Stacey A.
- Journal of Biological Chemistry, Vol. 287, Issue 11
Small Molecule Inhibitors of Clostridium difficile Toxin B-Induced Cellular Damage
journal, February 2015
- Tam, John; Beilhartz, Greg L.; Auger, Anick
- Chemistry & Biology, Vol. 22, Issue 2
A Kinetic Isotope Effect Study on the Hydrolysis Reactions of Methyl Xylopyranosides and Methyl 5-Thioxylopyranosides: Oxygen versus Sulfur Stabilization of Carbenium Ions
journal, October 2001
- Indurugalla, Deepani; Bennet, Andrew J.
- Journal of the American Chemical Society, Vol. 123, Issue 44
Clinical Practice Guidelines for Clostridium difficile Infection in Adults: 2010 Update by the Society for Healthcare Epidemiology of America (SHEA) and the Infectious Diseases Society of America (IDSA)
journal, May 2010
- Cohen, Stuart H.; Gerding, Dale N.; Johnson, Stuart
- Infection Control & Hospital Epidemiology, Vol. 31, Issue 05
Catalytic mechanism of enzymic glycosyl transfer
journal, November 1990
- Sinnott, Michael L.
- Chemical Reviews, Vol. 90, Issue 7
Rho protein inactivation induced apoptosis of cultured human endothelial cells
journal, October 2002
- Hippenstiel, Stefan; Schmeck, Bernd; N'Guessan, Phillipe Dje
- American Journal of Physiology-Lung Cellular and Molecular Physiology, Vol. 283, Issue 4
PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010
- Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
Assay of proteins in the presence of interfering materials
journal, January 1976
- Bensadoun, André; Weinstein, David
- Analytical Biochemistry, Vol. 70, Issue 1
Clostridium difficile Toxins: Mechanism of Action and Role in Disease
journal, April 2005
- Voth, D. E.; Ballard, J. D.
- Clinical Microbiology Reviews, Vol. 18, Issue 2
Isofagomine Induced Stabilization of Glucocerebrosidase
journal, November 2008
- Kornhaber, Gregory J.; Tropak, Michael B.; Maegawa, Gustavo H.
- ChemBioChem, Vol. 9, Issue 16
The pharmacological chaperone isofagomine increases the activity of the Gaucher disease L444P mutant form of β-glucosidase: Effect of isofagomine on L444P acid β-glucosidase
journal, February 2010
- Khanna, Richie; Benjamin, Elfrida R.; Pellegrino, Lee
- FEBS Journal, Vol. 277, Issue 7
Glycosyltransferases: Structures, Functions, and Mechanisms
journal, June 2008
- Lairson, L. L.; Henrissat, B.; Davies, G. J.
- Annual Review of Biochemistry, Vol. 77, Issue 1
Oxygen-18 leaving group kinetic isotope effects on the hydrolysis of nitrophenyl glycosides. 2. Lysozyme and .beta.-glucosidase: acid and alkaline hydrolysis
journal, May 1981
- Rosenberg, Steven; Kirsch, Jack F.
- Biochemistry, Vol. 20, Issue 11
The low molecular mass GTP-binding protein Rho is affected by toxin A from Clostridium difficile.
journal, March 1995
- Just, I.; Selzer, J.; von Eichel-Streiber, C.
- Journal of Clinical Investigation, Vol. 95, Issue 3
Current State of Clostridium difficile Treatment Options
journal, July 2012
- Venugopal, A. A.; Johnson, S.
- Clinical Infectious Diseases, Vol. 55, Issue suppl 2
Mechanistic insights into glycosidase chemistry
journal, October 2008
- Vocadlo, David J.; Davies, Gideon J.
- Current Opinion in Chemical Biology, Vol. 12, Issue 5
Structure of the full-length Clostridium difficile toxin B
journal, July 2019
- Chen, Peng; Lam, Kwok-ho; Liu, Zheng
- Nature Structural & Molecular Biology, Vol. 26, Issue 8
Toxins A and B from Clostridium difficile differ with respect to enzymatic potencies, cellular substrate specificities, and surface binding to cultured cells.
journal, October 1997
- Chaves-Olarte, E.; Weidmann, M.; Eichel-Streiber, C.
- Journal of Clinical Investigation, Vol. 100, Issue 7
Guidelines for Diagnosis, Treatment, and Prevention of Clostridium difficile Infections
journal, January 2013
- Surawicz, Christina M.; Brandt, Lawrence J.; Binion, David G.
- American Journal of Gastroenterology, Vol. 108, Issue 4
Relationship between the inhibition constant (KI) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction
journal, December 1973
- Yung-Chi, Cheng; Prusoff, William H.
- Biochemical Pharmacology, Vol. 22, Issue 23
Importance of Toxin A, Toxin B, and CDT in Virulence of an Epidemic Clostridium difficile Strain
journal, August 2013
- Kuehne, Sarah A.; Collery, Mark M.; Kelly, Michelle L.
- The Journal of Infectious Diseases, Vol. 209, Issue 1
Enzymatic Transition States and Drug Design
journal, October 2018
- Schramm, Vern L.
- Chemical Reviews, Vol. 118, Issue 22
Phaser crystallographic software
journal, July 2007
- McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D.
- Journal of Applied Crystallography, Vol. 40, Issue 4