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Updated: May 24, 2026
Isolation and Characterization of Microvesicles from Peripheral Blood
Published on: January 6, 2017
Characterization of membrane-shed microvesicles from cytokine-stimulated β-cells using proteomics strategies
Giuseppe Palmisano1, Søren Skov Jensen, Marie-Catherine Le Bihan
1Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.
Abstract:
Microparticles and exosomes are two of the most well characterized membrane-derived microvesicles released either directly from the plasma membrane or released through the fusion of intracellular multivesicular bodies with the plasma membrane, respectively. They are thought to be involved in many significant biological processes such as cell to cell communication, rescue from apoptosis, and immunological responses. Here we report for the first time a quantitative study of proteins from β-cell-derived microvesicles generated after cytokine induced apoptosis using stable isotope labeled amino acids in cell culture combined with mass spectrometry. We identified and quantified a large number of β-cell-specific proteins and proteins previously described in microvesicles from other cell types in addition to new proteins located to these vesicles. In addition, we quantified specific sites of protein phosphorylation and N-linked sialylation in proteins associated with microvesicles from β-cells. Using pathway analysis software, we were able to map the most distinctive changes between microvesicles generated during growth and after cytokine stimulation to several cell death and cell signaling molecules including tumor necrosis factor receptor superfamily member 1A, tumor necrosis factor, α-induced protein 3, tumor necrosis factor-interacting kinase receptor-interacting serine-threonine kinase 1, and intercellular adhesion molecule 1.
Insights
This study quantifies proteins in beta-cell microvesicles during cytokine-induced apoptosis. It identifies beta-cell specific proteins and changes in signaling molecules, offering insights into cell death mechanisms.
Area of Science:
- Cell Biology
- Proteomics
- Biochemistry
Background:
- Microvesicles and exosomes are key mediators of intercellular communication.
- These vesicles play roles in apoptosis, immune responses, and cell survival.
- Understanding beta-cell derived microvesicles is crucial for metabolic research.
Purpose of the Study:
- To quantitatively analyze proteins in beta-cell microvesicles following cytokine-induced apoptosis.
- To identify beta-cell specific proteins and novel proteins within these microvesicles.
- To investigate protein modifications like phosphorylation and sialylation in response to apoptosis.
Main Methods:
- Stable isotope labeling by amino acids in cell culture (SILAC) combined with mass spectrometry.
- Quantitative proteomics to identify and quantify proteins in microvesicles.
- Pathway analysis to interpret changes in protein profiles.
Main Results:
- Identification and quantification of numerous beta-cell specific and novel proteins in microvesicles.
- Detection of specific sites of protein phosphorylation and N-linked sialylation.
- Distinct changes in cell death and signaling molecules, including TNF pathway members and ICAM1, were observed.
Conclusions:
- Cytokine-induced apoptosis significantly alters the proteomic cargo of beta-cell microvesicles.
- These microvesicles contain key signaling molecules involved in cell death pathways.
- The findings provide a quantitative proteomic landscape of beta-cell microvesicles during apoptosis.
