Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
Nat Methods. 2011 Jul 10;8(8):655-8. doi: 10.1038/nmeth.1647.
Despite progress in mass spectrometry (MS)-based phosphoproteomics, large-scale in vivo analyses remain challenging. Here we report a 'spike-in' stable-isotope labeling with amino acids in cell culture (SILAC) methodology using standards derived from labeled mouse liver cell lines, using which we analyzed insulin signaling. With this approach we identified 15,000 phosphosites and quantitatively compared 10,000 sites in response to insulin treatment, creating a very large, accurately quantified in vivo phosphoproteome dataset.
尽管基于质谱（MS）的磷酸化蛋白质组学取得了进展，但大规模的体内分析仍然具有挑战性。在这里，我们报告了一种使用源自标记的小鼠肝细胞系的标准品进行细胞培养中的“掺入”稳定同位素标记的氨基酸（SILAC）方法，我们使用该方法分析了胰岛素信号。通过这种方法，我们鉴定了 15000 个磷酸化位点，并定量比较了胰岛素处理后 10000 个位点，创建了一个非常大的、准确量化的体内磷酸化蛋白质组数据集。
