Cwp2 from <i>Clostridium difficile</i> exhibits an extended three domain fold and cell adhesion <i>in vitro</i>

Bradshaw, W.J.; Kirby, Jonathan M.; Roberts, April K.; Shone, Clifford C.; Acharya, K. Ravi

doi:10.1111/febs.14157

Cited by 23 publications

(24 citation statements)

References 46 publications

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“…Although CWPs involved in bacteria colonization have been identified, more CWPs need to be studied for their important roles in CDI and potential application in developing new therapeutics and prophylactics in C. difficile. Like other bacteria, C. difficile also possess multiple adhesins, and several CWPs have been previously characterized, including SlpA, Cwp2, Cwp6, CwpV, Cwp66, Cwp84 and Cwp19 (Waligora et al, 2001;Wright et al, 2005;Kirby et al, 2009;Dang et al, 2010;Reynolds et al, 2011;Bradshaw et al, 2017;Ferreira et al, 2017;Wydau-Dematteis et al, 2018). Although the exact role of each CWP in pathogenesis remains to be further elucidated, antibodies to many CWPs have been found in serum samples from CDI patients, and investigational vaccines targeting Cwp84 Sandolo et al, 2011) have been developed, indicating that certain CWPs are surface exposed in vivo and could be developed into vaccines (Pechine et al, 2005;Wright et al, 2008;Biazzo et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

“…The role of the C. difficile S-layer in colonization, immunity, virulence and viability has been well-studied (Calabi et al, 2002;Pechine et al, 2005;Ausiello et al, 2006;Pechine et al, 2007;Kirk et al, 2017). Several other CWPs such as Cwp84, Cwp2, Cwp66, and CwpV have been investigated, and play important roles in host cell adhesion and immune system evasion during CDI (Waligora et al, 2001;Emerson et al, 2009;Kirby et al, 2009;Bradshaw et al, 2017;Wydau-Dematteis et al, 2018). Kirby et al (Kirby et al, 2009) identified a surface-associated protease, Cwp84, which cleaves SlpA precursor into mature form.…”

Section: Introductionmentioning

confidence: 99%

“…Kirby et al (Kirby et al, 2009) identified a surface-associated protease, Cwp84, which cleaves SlpA precursor into mature form. Bradshaw et al (Bradshaw et al, 2017) determined the structure of Cwp2 and demonstrated that cwp2 mutation resulted in increased TcdA release and impaired cellular adherence in vitro. Cwp66, which works as an adhesin, and CwpV, which confers phase-variable phage infection resistance, were also characterized (Waligora et al, 2001;Emerson et al, 2009;Sekulovic et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Cwp22, a novel peptidoglycan cross‐linking enzyme, plays pleiotropic roles in Clostridioides difficile

Zhu

Bullock

2019

Environmental Microbiology

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Summary Clostridioides difficile is a Gram‐positive, spore‐forming, toxin‐producing anaerobe pathogen, and can induce nosocomial antibiotic‐associated intestinal disease. While production of toxin A (TcdA) and toxin B (TcdB) contribute to the main pathogenesis of C. difficile, adhesion and colonization of C. difficile in the host gut are prerequisites for disease onset. Previous cell wall proteins (CWPs) were identified that were implicated in C. difficile adhesion and colonization. In this study, we predicted and characterized Cwp22 (CDR20291_2601) from C. difficile R20291 to be involved in bacterial adhesion based on the Vaxign reverse vaccinology tool. The ClosTron‐generated cwp22 mutant showed decreased TcdA and TcdB production during early growth, and increased cell permeability and autolysis. Importantly, the cwp22 mutation impaired cellular adherence in vitro and decreased cytotoxicity and fitness over the parent strain in a mouse infection model. Furthermore, lactate dehydrogenase cytotoxicity assay, live‐dead cell staining and transmission electron microscopy confirmed the decreased cell viability of the cwp22 mutant. Thus, Cwp22 is involved in cell wall integrity and cell viability, which could affect most phenotypes of R20291. Our data suggest that Cwp22 is an attractive target for C. difficile infection therapeutics and prophylactics.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cwp22, a novel peptidoglycan cross‐linking enzyme, plays pleiotropic roles in Clostridioides difficile

Zhu

Bullock

2019

Environmental Microbiology

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“…The structure of the glycoside hydrolase domain of C. difficile S-layer-associated protein Cwp19 consisting of residues 28-388 has been determined to a high resolution. This is the sixth structure of the functional region of a protein from the S-layer of C. difficile to be determined after LMW SLP [40], Cwp84 [41,42], Cwp6 and Cwp8 [43] and Cwp2 [44]. We have identified a number of potential residues that are likely to be important active site residues and have partially characterised the activity of Cwp19-fr.…”

Section: Discussionmentioning

confidence: 99%

The molecular structure of the glycoside hydrolase domain of Cwp19 from Clostridium difficile

et al. 2017

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Clostridium difficile is a burden to healthcare systems around the world, causing tens of thousands of deaths annually. The S‐layer of the bacterium, a layer of protein found of the surface of cells, has received a significant amount of attention over the past two decades as a potential target to combat the growing threat presented by C. difficile infections. The S‐layer contains a wide range of proteins, each of which possesses three cell wall‐binding domains, while many also possess a “functional” region. Here, we present the high resolution structure of the functional region of one such protein, Cwp19 along with preliminary functional characterisation of the predicted glycoside hydrolase. Cwp19 has a TIM barrel fold and appears to possess a high degree of substrate selectivity. The protein also exhibits peptidoglycan hydrolase activity, an order of magnitude slower than that of lysozyme and is the first member of glycoside hydrolase‐like family 10 to be characterised. This research goes some way to understanding the role of Cwp19 in the S‐layer of C. difficile.DatabaseStructural data are available in the PDB under the accession numbers 5OQ2 and 5OQ3.

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“…a LMW SLP: 3CVZ (Fagan et al 2009 ). b Cwp2: 5NJL (Bradshaw et al 2017a ). c Cwp8: 5J7Q (Usenik et al 2017 ).…”

Section: Slpamentioning

confidence: 99%

The structure of the S-layer of Clostridium difficile

Bradshaw

Roberts

Shone

et al. 2017

J. Cell Commun. Signal.

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The nosocomially acquired pathogen Clostridium difficile is the primary causative agent of antibiotic associated diarrhoea and causes tens of thousands of deaths globally each year. C. difficile presents a paracrystalline protein array on the surface of the cell known as an S-layer. S-layers have been demonstrated to possess a wide range of important functions, which, combined with their inherent accessibility, makes them a promising drug target. The unusually complex S-layer of C. difficile is primarily comprised of the high- and low- molecular weight S-layer proteins, HMW SLP and LMW SLP, formed from the cleavage of the S-layer precursor protein, SlpA, but may also contain up to 28 SlpA paralogues. A model of how the S-layer functions as a whole is required if it is to be exploited in fighting the bacterium. Here, we provide a summary of what is known about the S-layer of C. difficile and each of the paralogues and, considering some of the domains present, suggest potential roles for them.

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Cwp2 from Clostridium difficile exhibits an extended three domain fold and cell adhesion in vitro

Cited by 23 publications

References 46 publications

Cwp22, a novel peptidoglycan cross‐linking enzyme, plays pleiotropic roles in Clostridioides difficile

Cwp22, a novel peptidoglycan cross‐linking enzyme, plays pleiotropic roles in Clostridioides difficile

The molecular structure of the glycoside hydrolase domain of Cwp19 from Clostridium difficile

The structure of the S-layer of Clostridium difficile

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