Tsai Chia-Feng, Ku Wei-Chi, Chen Yu-Ju, Ishihama Yasushi
Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan.
School of Medicine, College of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan.
Methods Mol Biol. 2017;1636:313-325. doi: 10.1007/978-1-4939-7154-1_20.
Direct measurement of site-specific phosphorylation stoichiometry can unambiguously distinguish whether the degree of phosphorylation is regulated by upstream kinase/phosphatase activity or by transcriptional regulation to alter protein expression level. Here, we describe a motif-targeting quantitative proteomic approach that integrates dephosphorylation, isotope tag labeling, and enzymatic kinase reaction for large-scale phosphorylation stoichiometry measurement of the human proteome.
特定位点磷酸化化学计量的直接测量能够明确区分磷酸化程度是受上游激酶/磷酸酶活性调控，还是受转录调控以改变蛋白质表达水平。在此，我们描述了一种基序靶向定量蛋白质组学方法，该方法整合了去磷酸化、同位素标签标记和酶促激酶反应，用于大规模测量人类蛋白质组的磷酸化化学计量。
