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J Proteome Res. 2019 Mar 1;18(3):947-959. doi: 10.1021/acs.jproteome.8b00647. Epub 2019 Jan 23.

Quantitative Proteomic Analysis of Small and Large Extracellular Vesicles (EVs) Reveals Enrichment of Adhesion Proteins in Small EVs.

Author information

1
Department of Cell and Developmental Biology , Vanderbilt University School of Medicine , Nashville , Tennessee 37232 , United States.
2
Department of Internal Medicine , University of New Mexico , Albuquerque , New Mexico 87131 , United States.
3
Sarah Cannon Research Institute , Nashville , Tennessee 37203 , United States.
4
Department of Medicine , Vanderbilt University Medical Center , Nashville , Tennessee 37212 , United States.
5
Department of Biological Sciences , Vanderbilt University School of Medicine , Nashville , Tennessee 37212 , United States.
6
Department of Biostatistics , Vanderbilt University Medical Center , Nashville , Tennessee 37232 , United States.
7
Department of Pathology, Microbiology and Immunology , Vanderbilt University Medical Center , Nashville , Tennessee 37212 , United States.

Abstract

Extracellular vesicles (EVs) are important mediators of cell-cell communication due to their cargo content of proteins, lipids, and RNAs. We previously reported that small EVs (SEVs) called exosomes promote directed and random cell motility, invasion, and serum-independent growth. In contrast, larger EVs (LEVs) were not active in those assays, but might have unique functional properties. In order to identify protein cargos that may contribute to different functions of SEVs and LEVs, we used isobaric tags for relative and absolute quantitation (iTRAQ)-liquid chromatography (LC) tandem mass spectrometry (MS) on EVs isolated from a colon cancer cell line. Bioinformatics analyses revealed that SEVs are enriched in proteins associated with cell-cell junctions, cell-matrix adhesion, exosome biogenesis machinery, and various signaling pathways. In contrast, LEVs are enriched in proteins associated with ribosome and RNA biogenesis, processing, and metabolism. Western blot analysis of EVs purified from two different cancer cell types confirmed the enrichment of cell-matrix and cell-cell adhesion proteins in SEVs. Consistent with those data, we found that cells exhibit enhanced adhesion to surfaces coated with SEVs compared to an equal protein concentration of LEVs. These data suggest that a major function of SEVs is to promote cellular adhesion.

KEYWORDS:

adhesion; exosomes; extracellular vesicles; iTRAQ; microvesicles; proteomics

PMID:
30608700
PMCID:
PMC6411421
[Available on 2020-03-01]
DOI:
10.1021/acs.jproteome.8b00647

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