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Characterization of Immune Cell-derived Extracellular Vesicles and Studying Functional Impact on Cell Environment
Published on: June 2, 2020
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Reference and Ghost Proteins Identification in Rat C6 Glioma Extracellular Vesicles
Adriana-Natalia Murgoci1, Tristan Cardon2, Soulaimane Aboulouard2
1Univ. Lille, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse-PRISM, Lille 59000, France; Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava 84510, Slovakia.
Iscience
|April 26, 2020
Summary
Extracellular vesicles (EVs) are key in cell communication. This study reveals that treating EVs with enzymes exposes hidden surface proteins and identifies novel "ghost proteins," potentially driving glioma tumor growth.
Area of Science:
- Cell Biology
- Biochemistry
- Oncology
Background:
- Extracellular vesicles (EVs) are crucial for intercellular communication and have therapeutic potential.
- Current methods for EV isolation may not fully remove protein aggregates or surface-associated proteins.
Purpose of the Study:
- To develop a method to separate isolated EVs from protein aggregates and external membrane proteins.
- To identify surface proteins demasked by enzymatic digestion and discover novel EV-associated proteins.
Main Methods:
- EVs pellets underwent Trypsin/Lys C digestion and Amicon filtration.
- Proteomic analyses were performed on all resulting fractions.
- Comparison of proteomic data between digested and non-digested EVs.
Main Results:
- Enzymatic digestion revealed quantitative variations in EV surface proteins.
- Specific proteins (e.g., CD81, CD82, Lamp 1) were demasked after 6-hour digestion.
- Six ghost proteins were identified, including one long noncoding RNA.
Conclusions:
- EVs contain ghost proteins, some of which may be uncovered by enzymatic treatment.
- These ghost proteins, identified in glioma-derived EVs, could play a role in tumorigenesis.
