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Proteomic dataset: Profiling of cultivated Echerichia coli isolates from Crohn's disease patients and healthy individuals.

ABSTRACT: One of the dysbioses often observed in Crohn's disease (CD) patients is an increased abundance of Escherichia coli (10-100 fold compared to healthy individuals) (Gevers et al., 2014). The data reported is a large-scale proteome profile for E. coli isolates collected from CD patients and healthy individuals. 43 isolates were achieved from 30 CD patients (17 male, 12 female, median age 30) and 19 isolates from 7 healthy individuals (7 male, median age 19). Isolates were cultivated on LB medium at aerobic conditions up to medium log phase. Protein extraction was performed with sodium deoxycholate (DCNa) and urea, alcylation with tris(2-carboxyethyl)phosphine and iodacetamide. Protein trypsinolysis was performed as described in (Matyushkina et al., 2016). Total cell proteomes were analysed by shotgun proteomics with HPLC-MS/MS on a maXis qTOF mass-spectrometer. The data including HPLC-MS/MS raw files and exported Mascot search results was deposited to the PRIDE repository project accession: PXD010920, project https://doi.org/10.6019/PXD010920.

SUBMITTER: Rakitina D

PROVIDER: S-EPMC6660468 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Proteomic dataset: Profiling of cultivated Echerichia coli isolates from Crohn's disease patients and healthy individuals.

Rakitina Daria D Baikova Julia J Pobeguts Olga O Bukato Olga O Butenko Ivan I Garanina Irina I Levites Mark M Manolov Alexander A Kanygina Alexandra A Kostryukova Elena E Semashko Tatiana T Karpova Irina I Babenko Vladislav V Scherbakov Petr P Khalif Igor I Shapina Marina M Parfenov Asfold A Ruchkina Irina I Knyazev Oleg O Fadeeva Nina N Subbotin Alexandr A Chamkin Sergey S Pyrkh Alexandr A Ivantsova Marina M Govorun Vadim V

Data in brief 20190307

One of the dysbioses often observed in Crohn's disease (CD) patients is an increased abundance of Escherichia coli (10-100 fold compared to healthy individuals) (Gevers et al., 2014). The data reported is a large-scale proteome profile for E. coli isolates collected from CD patients and healthy individuals. 43 isolates were achieved from 30 CD patients (17 male, 12 female, median age 30) and 19 isolates from 7 healthy individuals (7 male, median age 19). Isolates were cultivated on ...[more]

PMID: 31372401

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Project description:AimTo explore the diagnostic models of Crohn's disease (CD), Intestinal tuberculosis (ITB) and the differential diagnostic model between CD and ITB by analyzing serum proteome profiles.MethodsSerum proteome profiles from 30 CD patients, 21 ITB patients and 30 healthy controls (HCs) were analyzed by using weak cationic magnetic beads combined with MALDI-TOF-MS technique to detect the differentially expressed proteins of serum samples. Three groups were made and compared accordingly: group of CD patients and HCs, group of ITB patients and HCs, group of CD patients and ITB patients. Wilcoxon rank sum test was used to screen the ten most differentiated protein peaks (P < 0.05). Genetic algorithm combining with support vector machine (SVM) was utilized to establish the optimal diagnostic models for CD, ITB and the optimal differential diagnostic model between CD and ITB. The predictive effects of these models were evaluated by Leave one out (LOO) cross validation method.ResultsThere were 236 protein peaks differently expressed between group of CD patients and HCs, 305 protein peaks differently expressed between group of ITB patients and HCs, 332 protein peaks differently expressed between group of CD patients and ITB patients. Ten most differentially expressed peaks were screened out between three groups respectively (P < 0.05) to establish diagnostic models and differential diagnostic model. A diagnostic model comprising of four protein peaks (M/Z 4964, 3029, 2833, 2900) can well distinguish CD patients and HCs, with a specificity and sensitivity of 96.7% and 96.7% respectively. A diagnostic model comprising four protein peaks (M/Z 3030, 2105, 2545, 4210) can well distinguish ITB patients and HCs, with a specificity and sensitivity of 93.3% and 95.2% respectively. A differential diagnostic model comprising three potential biomarkers protein peaks (M/Z 4267, 4223, 1541) can well distinguish CD patients and ITB patients, with a specificity and sensitivity of 76.2% and 80.0% respectively. Among the eleven protein peaks from the diagnostic models and differential diagnostic model, two have been successfully purified and identified, Those two peaks were M/Z 2900 from the diagnostic model between CD and HCs and M/Z 1541 from the differential diagnostic model between CD and ITB. M/Z 2900 was identified as appetite peptide, M/Z 1541 was identified as Lysyl oxidase-like 2 (LOXL-2).ConclusionThe differently expressed protein peaks analyzed by serum proteome with weak cationic magnetic beads combined MALDI-TOF-MS technique can effectively distinguish CD patients and HCs, ITB patients and HCs, CD patients and ITB patients. The diagnostic model between CD patients and HCs consisting of four protein peaks (M/Z 4964, 3029, 2833, 2900), the diagnostic model between ITB patients and HCs comprising four protein peaks (M/Z 3030, 2105, 2545, 4210) and the differential diagnostic model between CD patients and ITB patients comprising three protein peaks (M/Z 4267, 4223, 1541) had high specificity and sensitivity and can contribute to diagnoses of CD, ITB and the differential diagnosis between CD and ITB. Two proteins from the diagnostic model of CD and the differential diagnostic model between CD and ITB were identified. Further experiments are required using a larger cohort of samples.

Project description:Bifidobacteria are frequently used in probiotic food and dairy products. Bifidobacterium bifidum S17 is a promising probiotic candidate strain that displays strong adhesion to intestinal epithelial cells and elicits potent anti-inflammatory capacity both in vitro and in murine models of colitis. The recently sequenced genome of B. bifidum S17 has a size of about 2.2 Mb and encodes 1,782 predicted protein-coding genes. In the present study, a comprehensive proteomic profiling was carried out to identify and characterize proteins expressed by B. bifidum S17. A total of 1148 proteins entries were identified by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), representing 64.4% of the predicted proteome. 719 proteins could be assigned to functional categories according to cluster of orthologous groups of proteins (COGs). The COG distribution of the detected proteins highly correlates with that of the complete predicted proteome suggesting a good coverage and representation of the genomic content of B. bifidum S17 by the proteome. COGs that were highly present in the proteome of B. bifidum S17 were Translation, Amino Acid Transport and Metabolism, and Carbohydrate Transport and Metabolism. Complete sets of enzymes for both the bifidus shunt and the Embden-Meyerh of pathway were identified. Further bioinformatic analysis yielded 28 proteins with a predicted extracellular localization including 14 proteins with an LPxTG-motif for cell wall anchoring and two proteins (elongation factor Tu and enolase) with a potential moonlighting function in adhesion. Amongst the predicted extracellular proteins were five of six pilin proteins encoded in the B. bifidum S17 genome as well as several other proteins with a potential role in interaction with host structures. The presented results are the first compilation of a proteomic reference profile for a B. bifidum strain and will facilitate analysis of the molecular mechanisms of physiology, host-interactions and beneficial effects of a potential probiotic strain.

Project description:Crohn’s disease (CD) is a severe chronic immune-mediated granulomatous inflammatory disease of the gastrointestinal tract. Its pathogenesis mechanisms remain obscure. One of the factors of CD pathogenesis is reduced diversity of gut microbiota which is accompanied by the characteristic increase of representation of Escherichia coli strains, several of which are able to penetrate the mucin layer, adhere to the epithelial cells, cross the epithelial barrier, and colonize macrophages. During the internalization, these E. coli strains can modulate macrophages’ defensive functions in a way that allows them to survive in a presence of such negative factors as low pH, oxidative stress, proteolytic enzymes and antimicrobial solutions. These strains were collectively assigned to the adhesive–invasive group of E. coli (AIEC). Earlier we sequenced 28 E. coli isolates from 10 patients with CD [Rakitina DV et al. Genome analysis of E. coli isolated from Crohn disease patients. BMC Genomics. 2017, 18(1):544. doi: 10.1186/s12864-017-3917-x] It has been demonstrated that the majority of enteropathogens feature specific metabolic pathways allowing them to use alternative carbon sources which leads to survivability and enhaced competitiveness in the host organism. Previously, we demonstrated that virulent properties of CD isolates are dependent on the carbon source: prolonged growth on propionate containing medium stimulates virulent properties while prolonged growth on glucose reduces these properties to levels indistinguishable from laboratory strain K-12 MG1655 [Pobeguts OV et al. Propionate Induces Virulent Properties of Crohn’s Disease-Associated Escherichia coli. Frontiers in Microbiology. 2020, 11:1460. doi: 10.3389/fmicb.2020.01460]. Thus, cultivation of E. coli on different growth mediums: M9 supplied with propionate and glucose made it possible to distinguish two states of each CD isolate: active (aAIEC) and inactive (iAIEC). In order to identify the major changes that occur upon change of carbon source, we carried out LC–MS-based proteomic analysis of aAIEC and iAIEC states of three CD isolates of E. coli ZvL2 (RCE07), BruB2 (RCE03) and К5 (RCE06) grown on glucose and propionate.

Dataset Information

Proteomic dataset: Profiling of cultivated Echerichia coli isolates from Crohn's disease patients and healthy individuals.

Publications

Proteomic dataset: Profiling of cultivated <i>Echerichia coli</i> isolates from Crohn's disease patients and healthy individuals.

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