Characterization of Membrane-shed Microvesicles from Cytokine-stimulated β-Cells Using Proteomics Strategies
Characterization of Membrane-shed Microvesicles from Cytokine-stimulated β-Cells Using Proteomics Strategies
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DOI：
10.1074/mcp.m111.012732
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发表时间：
2012-08-01
影响因子：
7
通讯作者：
Larsen, Martin Rossel
中科院分区：
文献类型：
--
作者：
Palmisano, Giuseppe;Jensen, Soren Skov;Larsen, Martin Rossel
微颗粒和外泌体是两种最具特征的膜源性微泡，它们可以直接从质膜释放，也可以通过细胞内多泡体与质膜融合释放。它们被认为参与了许多重要的生物学过程，如细胞间通讯、细胞凋亡的拯救和免疫反应。在这里，我们首次报道了利用细胞培养中稳定同位素标记的氨基酸结合质谱法对细胞因子诱导凋亡后产生的β细胞来源的微泡中的蛋白质进行定量研究。我们鉴定并量化了大量的β细胞特异性蛋白质和先前在其他细胞类型的微泡中描述的蛋白质，以及位于这些微泡上的新蛋白质。此外，我们量化了β细胞中与微囊泡相关的蛋白质磷酸化和n -连接sia- lyylation的特定位点。利用通路分析软件，我们能够绘制出生长过程中和细胞因子刺激后产生的微泡与几种细胞死亡和细胞信号分子之间最显著的变化，包括肿瘤坏死因子受体超家族成员1A、肿瘤坏死因子、α诱导蛋白3、肿瘤坏死因子相互作用激酶受体相互作用丝氨酸-苏氨酸激酶1和细胞间粘附分子1。中国生物医学工程学报（自然科学版），2011,32(2):444 - 444。
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 beta-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 beta-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 sia-lylation in proteins associated with microvesicles from beta-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, alpha-induced protein 3, tumor necrosis factor-interacting kinase receptor-interacting serine-threonine kinase 1, and intercellular adhesion molecule 1. Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.012732, 230-243, 2012.