Functional Properties of Circulating Exosomes Mediated by Surface-Attached Plasma Proteins

Tatiana Shtam, Stanislav Naryzhny, Arthur Kopylov, Elena Petrenko, Roman Samsonov, Roman Kamyshinsky, Yana Zabrodskaya, Daniil Nikitin, Maxim Sorokin, Anton Buzdin, Anastasia Malek

Abstract


Background: Exosomes and other types of extracellular vesicles present an important component of circulating plasma. Exosomes released by endothelial and blood cells account for majority of plasma exosomal population, exosomes secreted by other cells might cross tissue-plasma barrier and reach circulating plasma as well. Definitely, exosomes of different cellular origin are different by content and function. However, exosomal surface membrane interacts with plasma components. This interaction may alter composition of exosomal surface and, hence, provide these vesicles with new functional properties.

Aims: This study was aimed to estimate composition and possible functional role of proteins attached on the surface of plasma exosomes.

Methods: Here, extracellular vesicles from human plasma were isolated by ultracentrifugation and treated by trypsin. Trypsinized and native exosomes were analyzed by nanoparticle tracking analysis, western blotting, and quantitative high-resolution mass spectrometry.

Results: Surface-attached proteins were removed from exosomes isolated from plasma of healthy female donors by incubation with serine protease (trypsin). Treatment did not impact exosomes integrity while slightly reduced hydrodynamic radius. Mass spectrometry revealed 261 exosomal proteins, among them 71 proteins were completely removed and more than half of the proteins were partially removed by trypsinization. Gene ontology functional annotation revealed mostly extracellular locations of proteins cleaved from a surface of the plasma exosomes. Moreover, proteins cleaved from the exosome surface are supposed to be implicated into Integrin –linked kinase (ILK), focal adhesion kinase (FAK) and other pathways connecting cell surface with intracellular signaling cascades.

Conclusions: Taken together, our results demonstrate that a surface of circulating exosomes is decorated by plasma proteins, and these proteins (1) can mask tissue-specific characteristic of the exosomal surface membrane and (2) provide exosomes with new and uniform properties.




J Hematol. 2018;7(4):149-153
doi: https://doi.org/10.14740/jh412w

Keywords


Exosomes; Plasma proteins; Mass spectrometry; FAK signaling; ILK signaling

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Journal of Hematology, quarterly, ISSN 1927-1212 (print), 1927-1220 (online), published by Elmer Press Inc.               
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