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Chemical approaches for the identification and quantitative analysis of protein phosphorylation by mass spectrometry

机译:质谱法鉴定和定量分析蛋白质磷酸化的化学方法

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Protein phosphorylation plays a central role in the regulation of many cellular processes. A number of mass spectrometric approaches have been used to identify protein phosphorylation sites, but the rapid identification of candidate phosphopeptides in single-stage mass spectrometry (MS) has been difficult, due to the frequently reduced representation of phosphopeptide signals in MS spectra. Furthermore, site-specific quantitative measurement of phosphorylation remains an outstanding challenge. Here we describe two related chemical approaches for the identification and quantitative analysis of protein phosphorylation by MS. Both approaches rely on in-gel chemical dephosphorylation to "uncover" phosphopeptides in the sample and have been used to study substrates of Aurora B, a kinase required for bipolar spindle formation in mitosis.
机译:蛋白质磷酸化在许多细胞过程的调节中起着核心作用。已经使用许多质谱方法来鉴定蛋白质磷酸化位点,但由于MS光谱中的磷肽信号的常量表示,难以识别单级质谱(MS)中的候选磷酸肽的快速鉴定。此外,特异性特异性磷酸化的定量测量仍然是一个突出的挑战。在这里,我们描述了两种相关化学方法,用于鉴定和定量分析MS蛋白质磷酸化。两种方法依赖于凝胶化学磷酸化物中的“揭开”样品中的磷酸肽,并已用于研究Aurora B的底物,在有丝分裂中进行双极主轴形成所需的激酶。

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