Dataset Information

Lactobacillus mucosae LM1 and intestinal cell interaction

ABSTRACT: In this study, we employed a gel-free proteomic approach to investigate proteome changes in both LM1 and IPEC-J2 cells after co-incubation. QExactive Orbitrap mass spectrometry (MS) was used for a full proteomic scan of bacterial and intestinal cells, and for a large-scale quantitative analysis of protein dynamics during host-microbe interaction. This is the first proteomic study to use this method to obtain novel insights into probiotic adhesion to IECs.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Lactobacillus Mucosae Lm1 Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: Edward Pajarillo

LAB HEAD: Dae-Kyung Kang

PROVIDER: PXD006851 | Pride | 2018-10-24

REPOSITORIES: Pride

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Publications

Proteomic View of the Crosstalk between Lactobacillus mucosae and Intestinal Epithelial Cells in Co-culture Revealed by Q Exactive-Based Quantitative Proteomics.

Pajarillo Edward Alain B EAB Kim Sang Hoon SH Valeriano Valerie Diane VD Lee Ji Yoon JY Kang Dae-Kyung DK

Frontiers in microbiology 20171212

Lactobacilli are bacteria that are beneficial to host health, but information on communication between Lactobacilli and host cells in the intestine is lacking. In this study, we examined the proteomes of the Lactobacillus mucosae strain LM1, as a model of beneficial bacteria, and the intestinal porcine epithelial cell line (IPEC-J2) after co-culture. Label-free proteomics demonstrated the high-throughput capability of the technique, and robust characterization of the functio ...[more]

PMID: 29312173

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Project description:Medium chain fatty acids (MCFA) have been shown to inhibit methanogenesis, disrupt the cell envelope, and decrease survival of Methanobrevibacter ruminantium M1 in a dose-, time-, and protonation level dependent way. However, the exact mechanisms behind these observations are still unknown. Although the biochemistry of the metabolic processes of M. ruminantium has been well studied and its genome sequence is now available, little is known about the overall transcriptome regulation of M. ruminantium in response to inhibitors like MCFA. In the present study, we used RNA Sequencing to evaluate the effects of lauric acid (C12) on M. ruminantium. Pure M. ruminantium cell cultures in the mid-exponential growth phase were exposed to C12 in concentrations of 0.4 mg/mL, dissolved in DMSO for 1 h, and the transcriptomic changes compared to DMSO-only treated control samples (final DMSO concentration 0.2 %), were investigated. Gene expression changes upon exposure to C12 were not dramatic in magnitude (log2 fold change mostly below +/- 3) and in gene number (214 genes). However, the observed expression changes affected mostly genes which encoded cell-surface associated proteins (adhesion-like proteins, membrane-associated transporters and hydrogenases), or proteins, which were involved in detoxification or DNA repair processes. The transcriptional response of M. ruminantium M1 to C12 did not specifically inhibit methanogenesis. Instead, the data indicated a non-specific antimicrobial action by lauric acid, which involves destruction of the cell membrane and interferences with cellular energetics in M. ruminantium. To date, there has been no systematic characterization of a ruminal methanogen transcriptome by deep sequencing. Our results give first hints on the molecular inhibitory mechanism of C12 on M. ruminantium. RNA profiles of M. ruminantium treated in vitro (lauric acid disolved in DMSO), non-treated=controls (DMSO supplementation) and non-treated=blanks (no supplementation) were generated by deep sequencing, in triplicates, by using Illumina HiSeq2500

Dataset Information

Lactobacillus mucosae LM1 and intestinal cell interaction

Publications

Proteomic View of the Crosstalk between <i>Lactobacillus mucosae</i> and Intestinal Epithelial Cells in Co-culture Revealed by Q Exactive-Based Quantitative Proteomics.

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