EspP, a Type V-secreted serine protease of enterohaemorrhagic<i>Escherichia coli</i>O157:H7, influences intestinal colonization of calves and adherence to bovine primary intestinal epithelial cells

Dziva, Francis; Mahajan, Aditya; Cameron, Pamela; Currie, Colin; McKendrick, Iain J.; Wallis, Timothy S.; Smith, David George Emslie; Stevens, Mark P.

doi:10.1111/j.1574-6968.2007.00724.x

Cited by 74 publications

(60 citation statements)

References 46 publications

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“…In agreement with a previous report suggesting that EspP is required for intestinal colonization in calves (20), we found that EspP interacts with fibronectin, indicating that EspP possesses lectin properties (Fig. 5C) and may recognize host proteins.…”

Section: Biochemical and Physical Properties Of The E Coli Ropes-cd supporting

confidence: 81%

“…EspC and EspP display enzymatic activities, and although EspC cleaves spectrin (also called fodrin) and hemoglobin, EspP of EHEC cleaves pepsin A, and human coagulation factor V (2,11,14,(17)(18)(19). The EspP protein was also shown to influence gut colonization in calves and contributes to adherence of EHEC to bovine primary rectal cells (20). Recently, EspP was found to be directly involved in biofilm formation and adherence of EHEC to T84 intestinal epithelial cells (21).…”

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confidence: 99%

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Bacterial Macroscopic Rope-like Fibers with Cytopathic and Adhesive Properties

Xicohtencatl‐Cortes

Saldaña

Deng

et al. 2010

Journal of Biological Chemistry

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We present a body of ultrastructural, biochemical, and genetic evidence that demonstrates the oligomerization of virulence-associated autotransporter proteins EspC or EspP produced by deadly human pathogens enterohemorrhagic and enteropathogenic Escherichia coli into novel macroscopic ropelike structures (>1 cm long). The rope-like structures showed high aggregation and insolubility, stability to anionic detergents and high temperature, and binding to Congo Red and thioflavin T dyes. These are properties also exhibited by human amyloidogenic proteins. These macroscopic ropes were not observed in cultures of nonpathogenic Escherichia coli or isogenic espP or espC deletion mutants of enterohemorrhagic or enteropathogenic Escherichia coli but were produced by an Escherichia coli K-12 strain carrying a plasmid expressing espP. Purified recombinant EspP monomers were able to self-assemble into macroscopic ropes upon incubation, suggesting that no other protein was required for assembly. The ropes bound to and showed cytopathic effects on cultured epithelial cells, served as a substratum for bacterial adherence and biofilm formation, and protected bacteria from antimicrobial compounds. We hypothesize that these ropes play a biologically significant role in the survival and pathogenic scheme of these organisms.

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Section: Biochemical and Physical Properties Of The E Coli Ropes-cd supporting

confidence: 81%

mentioning

confidence: 99%

Bacterial Macroscopic Rope-like Fibers with Cytopathic and Adhesive Properties

Xicohtencatl‐Cortes

Saldaña

Deng

et al. 2010

Journal of Biological Chemistry

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“…From the changes observed in the protein content of epithelium and mucus following larval challenge it is apparent that the in vitro model provided a technique which can be usefully deployed in investigations into recently established or immediate tissue responses to larval challenge. The use of the described enzymatic methodology for the isolation of epithelial cells from the gastric mucosa was based on the previously described method for the isolation of gastric crypts of bovine mucosa described by Dziva et al [4]. Cytospins confirmed that there was minimal contamination from non-epithelial lamina propria derived cells, compared to other methods used to isolate epithelial cells, such as mucosal scrapings and the use of EDTA (preliminary data, not included here), and only minimal contamination with albumin was observed in 2-D gels.…”

Section: Discussionmentioning

confidence: 99%

“…The aim of the enzymatic dispersion of the epithelial cells was two-fold: (i) to release the larvae that were closely associated to the gastric mucosa, and (ii) to achieve dispersion of epithelial cells for the proteomic analysis. The enzymatic dispersion was based on the method described by Dziva et al [4].…”

Section: In Vitro Model and Sample Preparationmentioning

confidence: 99%

Proteomic approach to identify candidate effector molecules during the in vitro immune exclusion of infectiveTeladorsagia circumcinctain the abomasum of sheep

Athanasiadou

Pemberton²,

Jackson³

et al. 2008

Vet. Res.

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-In the present study we have employed an in vitro organ challenge model to study the post-challenge responses in parasite naïve and immune gastric tissue of sheep, in an attempt to identify the host derived factors involved in immune exclusion of Teladorsagia circumcincta larvae. Proteins present in the epithelial cells and mucus from ovine abomasa following parasite challenge in previously naïve and immune animals were analysed through Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-Tof)-MS and shotgun proteomics. MALDI-ToF analysis of epithelial cell lysates revealed that a number of proteins identified were differentially expressed in naïve and immune cells. These included intelectin and lysozymes, which were present at higher levels in epithelial cell lysates derived from immune samples. A large number of proteins were identified in the mucosal wash from immune tissue which were not present in the mucosal wash of the naïve tissue. Some of these proteins were present in washes of immune tissue prior to the parasite challenge including immunoglobulin A, galectin 14 and 15 and sheep mast cell protease 1. However, other proteins, such as calcium activated chloride channel and intelectin were only detected in the washings from the challenged tissue. The latter may be related to an enhanced mucus release, which may result in entrapment of infective larvae and thus reduced establishment in tissue that has been previously challenged with the parasite. In conclusion, several proteins have been identified which may be involved, either directly or indirectly, in the exclusion and immune elimination of incoming infective larvae. In the present study, the usefulness of the in vitro model has been confirmed, and the global proteomic approach has identified proteins that had not previously been associated with parasite exclusion from abomasal mucosa, such as the calcium activated chloride channel.immunity / mucus / proteomics / sheep / Teladorsagia circumcincta

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“…What drives the selection of particular genes to create a STEC pathogen is unknown. However, because the existence of a primarily bovine animal reservoir of infection is a major difference between STEC and other pathotypes of E. coli, some genes, such as ehxA and espP, may be acquired by STEC to facilitate survival and persistence in the bovine gut (37,38). Therefore, although some determinants may not be considered essential virulence factors for human infection, they may confer an advantage to STEC survival and transmission in a different environment, such as an animal reservoir of infection.…”

Section: Discussionmentioning

confidence: 99%

Shiga Toxin–producingEscherichia coliStrains Negative for Locus of Enterocyte Effacement

Newton¹,

Sloan²,

Bulach³

et al. 2009

Emerg. Infect. Dis.

100

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Most Shiga toxin-producing Escherichia coli (STEC) infections that are associated with severe sequelae such as hemolytic uremic syndrome (HUS) are caused by attaching and effacing pathogens that carry the locus of enterocyte effacement (LEE). However, a proportion of STEC isolates that do not carry LEE have been associated with HUS. To clarify the emergence of LEE-negative STEC, we compared the genetic composition of the virulence plasmids pO113 and pO157 from LEE-negative and LEE-positive STEC, respectively. The complete nucleotide sequence of pO113 showed that several plasmid genes were shared by STEC O157:H7. In addition, allelic profi ling of the ehxA gene demonstrated that pO113 belongs to a different evolutionary lineage than pO157 and that the virulence plasmids of LEEnegative STEC strains were highly related. In contrast, multilocus sequence typing of 17 LEE-negative STEC isolates showed several clonal groups, suggesting that pathogenic LEE-negative STEC has emerged several times throughout its evolution.

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EspP, a Type V-secreted serine protease of enterohaemorrhagicEscherichia coliO157:H7, influences intestinal colonization of calves and adherence to bovine primary intestinal epithelial cells

Cited by 74 publications

References 46 publications

Bacterial Macroscopic Rope-like Fibers with Cytopathic and Adhesive Properties

Bacterial Macroscopic Rope-like Fibers with Cytopathic and Adhesive Properties

Proteomic approach to identify candidate effector molecules during the in vitro immune exclusion of infectiveTeladorsagia circumcinctain the abomasum of sheep

Shiga Toxin–producingEscherichia coliStrains Negative for Locus of Enterocyte Effacement

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