Dissection of the insulin signaling pathway via quantitative phosphoproteomics
Dissection of the insulin signaling pathway via quantitative phosphoproteomics
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DOI：
10.1073/pnas.0711713105
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发表时间：
2008-02-19
影响因子：
11.1
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Krueger, Marcus;Kratchmarova, Irina;Mann, Matthias
胰岛素信号传导途径在代谢疾病（如糖尿病）和细胞过程（如衰老）中具有关键重要性。胰岛素激活酪氨酸磷酸化级联反应，该级联反应分支形成影响多种生物过程的复杂网络。为了了解酪氨酸磷酸化级联反应的全谱，我们已经定义了胰岛素信号通路的酪氨酸磷酸化蛋白质组，使用高分辨率质谱结合磷酸酪氨酸免疫沉淀和稳定同位素标记细胞培养物中的氨基酸（SILAC）在分化的棕色脂肪细胞。在40种已鉴定的胰岛素诱导效应物中，有7种以前未在胰岛素信号传导中描述，包括SDR、PKC δ结合蛋白、LRP-6和PISP/PDZK 11（一种潜在的钙ATP酶结合蛋白）。使用PISP/PDZK 11的蛋白质组相互作用筛选鉴定了钙转运ATP酶SERCA 2，支持与钙信号传导的连接。定量磷酸化蛋白质组学与细胞培养模型的结合提供了一个强有力的策略来剖析完整细胞中的胰岛素信号通路。
The insulin signaling pathway is of pivotal importance in metabolic diseases, such as diabetes, and in cellular processes, such as aging. Insulin activates a tyrosine phosphorylation cascade that branches to create a complex network affecting multiple biological processes. To understand the full spectrum of the tyrosine phosphorylation cascade, we have defined the tyrosine-phosphoproteome of the insulin signaling pathway, using high resolution mass spectrometry in combination with phosphotyrosine immunoprecipitation and stable isotope labeling by amino acids in cell culture (SILAC) in differentiated brown adipocytes. Of 40 identified insulin-induced effectors, 7 have not previously been described in insulin signaling, including SDR, PKC delta binding protein, LRP-6, and PISP/PDZK11, a potential calcium ATPase binding protein. A proteomic interaction screen with PISP/PDZK11 identified the calcium transporting ATPase SERCA2, supporting a connection to calcium signaling. The combination of quantitative phosphoproteomics with cell culture models provides a powerful strategy to dissect the insulin signaling pathways in intact cells.