Proteomic profile of extracellular vesicles released by Lactiplantibacillus plantarum BGAN8 and their internalization by non-polarized HT29 cell line

• Published in: Scientific reports Vol. 10; no. 1; p. 21829
• Main Authors: Bajic, Svetlana Sokovic, Cañas, Maria-Alexandra, Tolinacki, Maja, Badia, Josefa, Sánchez, Borja, Golic, Natasa, Margolles, Abelardo, Baldomá, Laura, Ruas-Madiedo, Patricia
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 11-12-2020; Nature Publishing Group; Nature Portfolio
• Subjects: 631/326/41/1969; 631/326/41/2533; 631/326/41/2536; Amino acids; Bacteria; Bacterial Proteins - metabolism; Chlorpromazine; Clathrin; Cryoelectron Microscopy; Endocytosis; Epithelial cells; Exopolysaccharides; Extracellular vesicles; Extracellular Vesicles - metabolism; Extracellular Vesicles - ultrastructure; Glycolysis; Gram-positive bacteria; HT29 Cells; Humanities and Social Sciences; Humans; Internalization; Intestine; Lactobacillus plantarum - metabolism; Lipid rafts; Metabolic pathways; multidisciplinary; Nystatin; Proteomics; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-78920-z

In recent years the role of extracellular vesicles (EVs) of Gram-positive bacteria in host-microbe cross-talk has become increasingly appreciated, although the knowledge of their biogenesis, release and host-uptake is still limited. The aim of this study was to characterize the EVs released by the dairy isolate
Lactiplantibacillus plantarum
BGAN8 and to gain an insight into the putative mechanism of EVs uptake by intestinal epithelial cells. The cryo-TEM observation undoubtedly demonstrated the release of EVs (20 to 140 nm) from the surface of BGAN8, with exopolysaccharides seems to be part of EVs surface. The proteomic analysis revealed that the EVs are enriched in enzymes involved in central metabolic pathways, such as glycolysis, and in membrane components with the most abundant proteins belonging to amino acid/peptide ABC transporters. Putative internalization pathways were evaluated in time-course internalization experiments with non-polarized HT29 cells in the presence of inhibitors of endocytic pathways: chlorpromazine and dynasore (inhibitors of clathrin-mediated endocytosis—CME) and filipin III and nystatin (disrupting lipid rafts). For the first time, our results revealed that the internalization was specifically inhibited by dynasore and chlorpromazine but not by filipin III and nystatin implying that one of the entries of
L. plantarum
vesicles was through CME pathway.