title: Characterization of membrane-shed microvesicles from cytokine-stimulated β-cells using proteomics strategies.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Palmisano
authors.forename: Søren Skov
authors.initials: SS
authors.lastname: Jensen
authors.forename: Marie-Catherine
authors.initials: MC
authors.lastname: Le Bihan
authors.forename: Jeanne
authors.initials: J
authors.lastname: Lainé
authors.forename: James N
authors.initials: JN
authors.lastname: McGuire
authors.forename: Flemming
authors.initials: F
authors.lastname: Pociot
authors.forename: Martin Røssel
authors.initials: MR
authors.lastname: Larsen
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002247:Carbon Isotopes; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D055252:Cell-Derived Microparticles / Q000378:metabolism* / Q000648:ultrastructure; D002853:Chromatography, Liquid; D016207:Cytokines / Q000494:pharmacology*; D055354:Exosomes / Q000378:metabolism* / Q000648:ultrastructure; D006023:Glycoproteins / Q000032:analysis; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D007371:Interferon-gamma / Q000494:pharmacology; D053583:Interleukin-1beta / Q000494:pharmacology; D007553:Isotope Labeling / Q000379:methods; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D009587:Nitrogen Isotopes; D010750:Phosphoproteins / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D051381:Rats; D053719:Tandem Mass Spectrometry; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes; D016207:Cytokines; D006023:Glycoproteins; D053583:Interleukin-1beta; D009587:Nitrogen Isotopes; D010750:Phosphoproteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma