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Characterization of membrane-shed microvesicles from cytokine-stimulated β-cells using proteomics strategies

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  • Giuseppe Palmisano
  • Søren Skov Jensen
  • Marie-Catherine Le Bihan
  • Jeanne Lainé
  • James N McGuire
  • Flemming Pociot
  • Martin Røssel Larsen
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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.
Original languageEnglish
JournalMolecular and Cellular Proteomics
Volume11
Issue number8
Pages (from-to)230-43
Number of pages14
ISSN1535-9476
DOIs
Publication statusPublished - Aug 2012

    Research areas

  • Amino Acids, Animals, Apoptosis, Carbon Isotopes, Cell Line, Tumor, Cell Membrane, Cell-Derived Microparticles, Chromatography, Liquid, Cytokines, Exosomes, Glycoproteins, Insulin-Secreting Cells, Interferon-gamma, Interleukin-1beta, Isotope Labeling, Microscopy, Electron, Transmission, Microscopy, Fluorescence, Nitrogen Isotopes, Phosphoproteins, Proteomics, Rats, Tandem Mass Spectrometry, Tumor Necrosis Factor-alpha

ID: 38661160