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J Proteomics. 2019 Feb 10;192:78-88. doi: 10.1016/j.jprot.2018.08.010. Epub 2018 Aug 25.

Identification of protein markers for extracellular vesicle (EV) subsets in cow's milk.

Author information

1
CHU de Québec Research Center/CHUL Pavilion, 2705 Blvd Laurier, Quebec City, QC, G1V 4G2 and Department of Microbiology, Infectious Diseases and Immunology, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada.
2
Proteomics Platform, Genomics Center, CHU de Québec Research Center/CHUL Pavilion, 2705 Blvd Laurier, Quebec City, QC G1V 4G2, Canada.
3
CHU de Québec Research Center/CHUL Pavilion, 2705 Blvd Laurier, Quebec City, QC, G1V 4G2 and Department of Microbiology, Infectious Diseases and Immunology, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada. Electronic address: patrick.provost@crchudequebec.ulaval.ca.

Abstract

Extracellular vesicles (EVs), like exosomes, are small membrane vesicles involved in cell-to-cell communications that modulate numerous biological processes. We previously discovered a new EV subset in milk (sedimenting at 35,000 g; 35 K) that protected its cargo (RNAs and proteins) during simulated digestion and was more enriched in microRNAs than exosomes (sedimenting at 100 K). Here, we used LC-MS/MS to push further the comparison between these two pellets. Commonly used EV markers were not differentially enriched between the pellets, questioning their use with cow's milk EVs. Similarly, the majority of the quantified proteins were equally enriched between the two pellets. Nevertheless, 20 proteins were specific to 35 K, while 41 were specifically enriched in 100 K (p < 0.05), suggesting their potential use as specific markers. Loaded with these proteins, the EVs in these pellets might regulate translation, proliferation and cell survival for 35 K, and metabolism, extracellular matrix turnover and immunity for 100 K. This approach also brought new insights into milk EV-associated integrins and their possible role in specifically targeting recipient cell types. These findings may help better discriminate between milk EVs, improve our understanding of milk EV-associated protein function and their possible use as therapeutic tools for the management of immunity- and metabolism-associated disorders. WEB PAGE: http://www.crchuq.ca/en/research/researchers/4691.

KEYWORDS:

Cow milk; Exosomes; Extracellular vesicles; Mass-spectrometry; Protein markers; Tetraspanins

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