Isolation and Characterization of Serum Extracellular Vesicles (EVs) from Atlantic Salmon Infected with Piscirickettsia Salmonis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacteria, Media, and Growth Conditions
2.2. Fish Husbandry and Challenge
2.3. Serum Exosome Isolation
2.4. Nanoparticle Tracking Analysis
2.5. Flow Cytometry
2.6. Transmission Electron Microscopy (TEM)
2.7. In-Solution Digestion and Protein Sequence Analysis by LC-MS/MS
2.8. Proteomic Data Analysis
2.9. Ethics Statement
3. Results
3.1. Serum EV Characterization
3.2. Serum EVs Proteome
4. Discussion
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Protein ID | Gen | Identified Proteins | Function |
---|---|---|---|
B5XFB2 | blvrb | Flavin reductase | Reductase activity |
B5X3Z5 | fgl2 | Fibroleukin | Peptidase activity |
Q9DDI8 | gdf-8 | Myostatin 1b | Receptor binding |
B5XF63 | natte | Nattectin | Carbohydrate binding |
C0H8M3 | tco2 | Transcobalamin-2 | Cobalamin binding |
B5X5M2 | fabph | Fatty acid-binding protein, heart | Transport activity |
B9EMN9 | leca | Lectin | Ion binding |
Protein | Identified Proteins | Function | Localization | Process |
---|---|---|---|---|
B5X3W8 | Galectin-3-binding protein | Receptor mediated endocytosis | membrane | Endocytosis |
Q9DD33 | Proteasome subunit beta type-9 | Proteolysis | cytoplasm | Immune system proccess |
B5X0V5 | Calreticulin | Calcium ion binding | endoplasmic reticulum | Protein folding |
C0HBQ7 | Caprin-1 | RNA binding | cytoplasmic | Regulation of translation |
A7KDZ9 | Proteasome subunit beta type | Proteolysis | cytoplasm | Proteosoma complex |
B5RI16 | Proteasome, 26S subunit, ATPase, 6 | Protein catabolic proccess | cytoplasm | ATP-binding |
B5X6D8 | Leukocyte cell-derived chemotaxin 2 | Protein binding | extracellular | Chemotaxis |
B5XF14 | Proteasome subunit alpha type | Protein catabolic proccess | cytoplasm | Endopeptidase activity |
C0PU67 | Gelsolin (Fragment) | Actin filament binding | cytoplasm | Actin nucleation |
Q49TU5 | Cathelicidin antimicrobial peptide | Protein binding | extracellular | Defense response to Gram-negative bacterium |
B5X0W4 | Eukaryotic translation initiation factor 3 subunit I | Translation | cytoplasm | Translation |
A7KE01 | Proteasome subunit beta type-6-A like protein | Endopeptidase activity | cytoplasm, nucleus | Immune system proccess |
B5RI28 | Proteasome, 26S subunit, ATPase 1a (Fragment) | Protein catabolic proccess | cytoplasm | ATP-binding |
B5X3K2 | Glyceraldehyde-3-phosphate dehydrogenase | Oxidoreductasa | cytoplasm | Glycolisis |
B5XAP5 | Protein S100 | Calcium binding | extracellular | Immune system process |
B5XDU3 | 14-3-3 protein zeta | Protein domain specific binding | cytoplasm | Signaling pathway |
C0H9S0 | Eukaryotic translation initiation factor 3 subunit A | Translation | cytoplasm | RNA binding |
C0PUP2 | Granulins (Fragment) | Protein binding | extracellular/exosome | Neutrophil degranulation |
B5X2B1 | Histone-binding protein RBBP4 | Protein binding | nucleus | Regulation of cell cycle |
A2VA22 | MHC class II antigen beta chain (Fragment) | Immune response | membrane | Antigen proccessing nad presentation |
C0PUI9 | Fructose-1,6-bisphosphatase 1 (Fragment) | Carbohydrate metabolic proccess | membrane | Hydrolase activity |
Q5UT54 | Toll-like leucine-rich repeat protein | LPS binding | secreted or membrane components | Immune response |
B5X141 | Ras-related protein Rab-10 | GTPase activity | cytoplasmic | GTP binding |
B5X1G4 | Cysteinyl-tRNA synthetase | Nucleotide binding | cytoplasmic | tRNA aminoacylation for protein translation |
B5X2E1 | Adenylyl cyclase-associated protein | Actin binding | membrane | Cytosqueleton organization |
B5X7T8 | Type-2 ice-structuring protein | Carbohydrate binding | secreted or membrane components | C-type lectin |
B5X8H5 | Peroxiredoxin | Antioxidant activity | cytoplasm | Cell redox homeostasis |
B5XDE4 | 14-3-3 protein beta/alpha | Protein domain specific binding | cytoplasm | Signaling pathway |
B9EQN9 | Plastin-2 | Calcium/actin binding | cytoplasm/cytosqueleton | Actin crosslink formation |
H8PHI0 | Cathelicidin-derived antimicrobial peptide 1 isoform A | LPS binding | cell | Defense response to Gram-negative bacterium |
B5X340 | Asparaginyl-tRNA synthetase, | Nucleotide binding | cytoplasmic | tRNA aminoacylation for protein translation |
B5X383 | SUMO-activating enzyme subunit 2 | Nucleotide/metal ion binding | cytoplasmic/nucleus | Protein sumoylation |
B5DGD5 | Actin related protein 2/3 complex subunit 2 | Actin binding | cytoesqueleton, extracellular exosome | Actin filament polymerization |
C0HAP0 | Class I histocompatibility antigen, F10 alpha chain | Antigen proccessing and presentation | membrane | Immune response |
B5X6E1 | Proteasome activator complex subunit 2 | Protein binding | proteasome complex/cytoplasm/nucleosome | Protein polyubiquitination |
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Lagos, L.; Tandberg, J.; Kashulin-Bekkelund, A.; Colquhoun, D.J.; Sørum, H.; Winther-Larsen, H.C. Isolation and Characterization of Serum Extracellular Vesicles (EVs) from Atlantic Salmon Infected with Piscirickettsia Salmonis. Proteomes 2017, 5, 34. https://doi.org/10.3390/proteomes5040034
Lagos L, Tandberg J, Kashulin-Bekkelund A, Colquhoun DJ, Sørum H, Winther-Larsen HC. Isolation and Characterization of Serum Extracellular Vesicles (EVs) from Atlantic Salmon Infected with Piscirickettsia Salmonis. Proteomes. 2017; 5(4):34. https://doi.org/10.3390/proteomes5040034
Chicago/Turabian StyleLagos, Leidy, Julia Tandberg, Alexander Kashulin-Bekkelund, Duncan J. Colquhoun, Henning Sørum, and Hanne C. Winther-Larsen. 2017. "Isolation and Characterization of Serum Extracellular Vesicles (EVs) from Atlantic Salmon Infected with Piscirickettsia Salmonis" Proteomes 5, no. 4: 34. https://doi.org/10.3390/proteomes5040034