Dataset Information

Proteomics of Diverse Clonal Groups of Enterotoxigenic Escherichia coli Underline Variations in Surface Antigens and Similarities in Metabolism

ABSTRACT: Enterotoxigenic Escherichia coli (ETEC) is an important cause of diarrhoea, a pervasive disease in low- and middle-income countries that affects mostly young children and visiting travellers. Substantial diversity exists among ETEC isolates, hindering development of effective preventive means. To investigate how ETEC genomic variation is reflected at expressed proteome level, we applied label-free quantitative proteomics to 7 human ETEC strains representing 5 epidemiologically important lineages. We further determined the protein profile of the non-pathogenic E. coli B strain BL21 to discriminate metabolic features specific for ETEC. The analysis yielded a dataset of 2,894 proteins, of which 1,777 were present in all strains. Each ETEC strain displayed on average 27 (±10) proteins with known or putative links to virulence, both plasmid- and chromosomally-encoded, and a number of strain-specific isoforms participating in the biosynthesis of surface antigens. Statistical comparison of relative protein levels between the ETEC strains and BL21 revealed several proteins with significantly increased amounts only in BL21, including enzymes of arginine biosynthesis and metabolism of melibiose, galactitol and gluconate. ETEC strains displayed consistently increased levels of proteins that were functional in iron acquisition, maltose metabolism, and acid resistance. These results suggest that specific metabolic functions are shared among ETEC isolates.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Escherichia Coli

DISEASE(S): Diarrhea

SUBMITTER: Veronika Kucharova

LAB HEAD: Harald G. Wiker

PROVIDER: PXD005259 | Pride | 2017-12-06

REPOSITORIES: Pride

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Publications

Comparative Proteomics of Enterotoxigenic Escherichia coli Reveals Differences in Surface Protein Production and Similarities in Metabolism.

Pettersen Veronika Kuchařová VK Steinsland Hans H Wiker Harald G HG

Journal of proteome research 20171212 1

Enterotoxigenic Escherichia coli (ETEC) infections are an important cause of diarrhea among young children living in low- and middle-income countries and visiting travelers. The development of effective vaccines is complicated by substantial genomic diversity that exists among ETEC isolates. To investigate how ETEC genomic variation is reflected at expressed proteome level, we applied label-free quantitative proteomics to seven human ETEC strains representing five epidemiologically important lin ...[more]

PMID: 29185342

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Project description:Enterotoxigenic Escherichia coli (ETEC) strains that produce both heat-stable (ST) and heat-labile (LT) enterotoxins cause severe post-weaning diarrhea in piglets. However, the relative importance of the individual enterotoxins to the pathogenesis of ETEC infection is poorly understood. In this study, we investigated the effect on virulence of an F4+ ETEC strain when removing some or all of its enterotoxins. Several isogenic mutant strains were constructed that lack the expression of LT in combination with one or both types of ST enterotoxins (STa and/or STb). Host early immune responses induced by these mutant strains 4h after infection were compared to the wild type strain GIS26 Â (O149:F4ac+, Â LT+ Â STa+ Â STb+). At the same time, the immune response of this wild type ETEC strain was compared to the mock-infected control, demonstrating the expression of porcine inflammatory response genes. For these purposes, the small intestinal segment perfusion (SISP) technique and microarray analysis were used and results were validated by qRT-PCR. We also measured net fluid absorption of pig small intestinal mucosa 4h after infection with wild type ETEC, the mutant strains and PBS (mock-infected). These data indicate an important role for STb in inducing small intestinal secretion early after infection. The microarray analysis of the different mutant strains also revealed an important role for STb in ETEC-induced immune response by the significant differential regulation of immune mediators like matrix metalloproteinase 3, interleukin 1 and interleukin 17. We conclude that STb can play a prominent role in ETEC-induced secretion and early immune response. In three pigs, 6 different treatments were performed. These treatments consisted of 4 mutant enterotoxigenic Escherichia coli GIS26 strains, GIS26 wild type strain, or PBS control. Per pig, the small intestine was divided into 6 loops with an interloop in between to avoid cross-contamination. In conclusion, every pig received each of the 6 treatments ad random.

			Action	DRS
	BL21_L1.raw	Raw
	BL21_L2.raw	Raw
	BL21_L4.raw	Raw
	BL21_T1.raw	Raw
	BL21_T2.raw	Raw