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Peptidomic and proteomic analysis of precision-cut lung slice supernatants

ABSTRACT: The data provided here is related to the publication “Peptidomic and proteomic analysis of precision-cut lung slice supernatants”. In this study, we sought out to develop a method that could incorporate a peptidomic and proteomic endpoint within the same workflow. The developed workflow was adapted from single-pot, solid-phase-enhanced sample preparation (SP3) for examining the proteome and peptidome of precision-cut lung slice supernatants in one experimental procedure to save sample material. A part of the study compared potential differences when utilizing different organic solvents for binding peptides and proteins to magnetic beads for biological replicates of PCLS supernatant. In connection, the workflow robustness was also characterized by investigating technical replicates of the same biological PCLS supernatant sample.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Lung

SUBMITTER: Lasse Gøbel Lorentzen

LAB HEAD: Micheal J

PROVIDER: PXD051333 | Pride | 2025-03-10

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	160101a_BE1_1_4205.pepXML	Pepxml
	160101a_BE1_1_4205_model.png	Other
	160101a_BE1_1_4205_quant.csv	Csv
	160101b_BE2_1_4206.pepXML	Pepxml
	160101b_BE2_1_4206_quant.csv	Csv

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Project description:Purpose: In this work, we evaluated the role of two indicative species, Citrobacter werkmanii (CW) and Escherichia albertii (EA), in the virulence of two DEC pathotypes, Shiga toxin-producing (STEC) and enteroaggregative (EAEC) Escherichia coli. Methods: To determine the effect of supernatant obtained from CW and EA cultures in STEC strain 86-24 and EAEC strain 042 gene expression, a RNA-seq analysis was performed. T84 cells were infected with DEC strains in the presence or absence of supernatant from EA and IL-8 secretion was evaluated. The effect of supernatant from EA on the growth and adherence of STEC and EAEC to T84 cells was also evaluated. Finally, we studied the participation of long polar fimbriae (Lpf) in STEC and plasmid-encoded toxin (Pet) in EAEC during DEC infection in the presence of supernatant from EA. Results: RNA-seq analysis revealed that several virulence factors in STEC and EAEC were up-regulated in the presence of supernatants from CW and EA. Interestingly, an increase in the secretion of IL-8 was observed in T84 cells infected with STEC or EAEC in the presence of a supernatant from EA. Similar results were observed with the supernatants obtained from clinical strains of E. albertii. Supernatant from EA had no effect on the growth of STEC and EAEC, or on the ability of these DEC strains to adhere to intestinal epithelial cells. Finally, we found that Pet toxin in EAEC was up-regulated in the presence of a supernatant from EA. In STEC, using mutant strains for Lpf fimbriae, our data suggested that these fimbriae might be participating in the increase of IL-8 induced by STEC on intestinal epithelial cells in the presence of a supernatant from EA. Conclusion:Supernatant obtained from an indicative species of DEC-positive diarrhea could modulate gene expression in STEC and EAEC, and IL-8 secretion induced by these bacteria. These data provide new insights into the effect of gut microbiota species in the pathogenicity of STEC and EAEC.

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Peptidomic and proteomic analysis of precision-cut lung slice supernatants

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