Cantin Greg T, Venable John D, Cociorva Daniel, Yates John R
Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Rd. SR11, La Jolla, California 92037, USA.
J Proteome Res. 2006 Jan;5(1):127-34. doi: 10.1021/pr050270m.
Protein phosphorylation has become a focus of many proteomic studies due to the central role that it plays in biology. We combine peptide-based gel-free isoelectric focusing and immobilized metal affinity chromatography to enhance the detection of phosphorylation events within complex protein samples using LC-MS. This method is then used to carry out a quantitative phosphoproteomic analysis of the tumor necrosis factor (TNF) pathway using HeLa cells metabolically labeled with 15N-containing amino acids, where 145 phosphorylation sites were found to be up-regulated upon the activation of the TNF pathway.
由于蛋白质磷酸化在生物学中发挥的核心作用，它已成为许多蛋白质组学研究的焦点。我们将基于肽的无胶等电聚焦和固定化金属亲和色谱相结合，以增强使用液相色谱 - 质谱法对复杂蛋白质样品中磷酸化事件的检测。然后，该方法用于对用含15N氨基酸进行代谢标记的HeLa细胞的肿瘤坏死因子（TNF）途径进行定量磷酸蛋白质组分析，结果发现有145个磷酸化位点在TNF途径激活后上调。
