Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed
Abstract
:1. Introduction
2. Results
2.1. Characterization of Vesicles Separated by Different Techniques
2.2. Proteomic Profile of EVs Enriched by Ultracentrifugation
2.3. Profile of Proteins Captured with a Precipitation Kit with Affinity for Glycosylated Groups
2.4. Differences between the Preparations Performed with Size-Exclusion-Based Method with Respect to the Polymer-Based Precipitation Method
3. Discussion
4. Materials and Methods
4.1. Obtention of Human Serum Samples
4.2. Enrichment of Extracellular Vesicles
4.3. EVs Characterization by Electron Microscopy and Nanoparticle-Tracking Analysis
4.4. Western Blot Analysis
4.5. Proteomics Analysis
4.6. Cell Component Enrichment Analysis and Information on Glycosylation Sites
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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#Term ID | Term Description | Term Size | Intersected | p Adjusted |
---|---|---|---|---|
GO:0022626 | cytosolic ribosome | 104 | 46 | 3.05 × 10−59 |
GO:0070062 | extracellular exosome | 2177 | 100 | 5.52 × 10−34 |
GO:0005925 | focal adhesion | 384 | 38 | 1.48 × 10−21 |
GO:0005739 | mitochondrion | 1380 | 66 | 5.04 × 10−21 |
GO:0015934 | large ribosomal subunit | 113 | 24 | 5.84 × 10−21 |
GO:0031966 | mitochondrial membrane | 573 | 44 | 7.98 × 10−21 |
GO:0072562 | blood microparticle | 138 | 44 | 2.97 × 10−79 |
GO:0070062 | extracellular exosome | 2177 | 51 | 4.92 × 10−34 |
GO:0062023 | collagen-containing extracellular matrix | 381 | 23 | 1.84 × 10−20 |
GO:0034358 | plasma lipoprotein particle | 23 | 7 | 2.93 × 10−10 |
GO:0031093 | platelet alpha granule lumen | 66 | 9 | 3.79 × 10−10 |
GO:0005579 | membrane attack complex | 6 | 5 | 7.24 × 10−10 |
GO:0042571 | immunoglobulin complex, circulating | 75 | 9 | 1.26 × 10−9 |
GO:0034361 | very-low-density lipoprotein particle | 15 | 6 | 1.91 × 10−9 |
GO:0034385 | triglyceride-rich plasma lipoprotein particle | 15 | 6 | 1.91 × 10−9 |
#Term ID | Term Description | Term Size | Intersected | p Adjusted |
---|---|---|---|---|
GO:0072562 | blood microparticle | 138 | 21 | 3.78 × 10−31 |
GO:1903561 | extracellular vesicle | 2262 | 32 | 1.37 × 10−17 |
GO:0031093 | platelet alpha granule lumen | 66 | 9 | 1.15 × 10−11 |
GO:0005579 | membrane attack complex | 6 | 4 | 1.15 × 10−7 |
#Term ID | Term Description | Term Size | Intersected | p Adjusted |
---|---|---|---|---|
GO:0072562 | blood microparticle | 138 | 20 | 1.13 × 10−32 |
GO:1990777 | lipoprotein particle | 23 | 8 | 5.31 × 10−15 |
GO:0070062 | extracellular exosome | 2177 | 25 | 8.04 × 10−15 |
GO:0042627 | chylomicron | 10 | 5 | 8.13 × 10−10 |
GO:0031089 | platelet dense granule lumen | 14 | 2 | 0.00270 |
GO:0070062 | extracellular exosome | 2177 | 7 | 0.00418 |
GO:0042788 | polysomal ribosome | 31 | 2 | 0.013765 |
#Term ID | Term Description | Term Size | Intersected | p Adjusted |
---|---|---|---|---|
GO:0072562 | blood microparticle | 138 | 22 | 1.47 × 10−38 |
GO:0070062 | extracellular exosome | 2177 | 21 | 1.16 × 10−10 |
GO:0071746 | IgA immunoglobulin complex, circulating | 5 | 3 | 1.03 × 10−5 |
GO:1990777 | lipoprotein particle | 23 | 4 | 1.39 × 10−5 |
GO:0031093 | platelet alpha granule lumen | 66 | 5 | 2.01 × 10−5 |
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Azkargorta, M.; Iloro, I.; Escobes, I.; Cabrera, D.; Falcon-Perez, J.M.; Elortza, F.; Royo, F. Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed. Int. J. Mol. Sci. 2021, 22, 11144. https://doi.org/10.3390/ijms222011144
Azkargorta M, Iloro I, Escobes I, Cabrera D, Falcon-Perez JM, Elortza F, Royo F. Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed. International Journal of Molecular Sciences. 2021; 22(20):11144. https://doi.org/10.3390/ijms222011144
Chicago/Turabian StyleAzkargorta, Mikel, Ibon Iloro, Iraide Escobes, Diana Cabrera, Juan M. Falcon-Perez, Felix Elortza, and Felix Royo. 2021. "Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed" International Journal of Molecular Sciences 22, no. 20: 11144. https://doi.org/10.3390/ijms222011144