Phosphorylation is arguably the most important post-translational modification that occurs within proteins. Phosphorylation is used as a signal to control numerous physiological activities ranging from gene expression to metabolism. Identifying phosphorylation sites within proteins was historically a challenge as it required either radioisotope labeling or the use of phospho-specific antibodies. The advent of mass spectrometry (MS) has had a major impact on the ability to qualitatively and quantitatively characterize phosphorylated proteins. In this article, we describe MS methods for characterizing phosphorylation sites within individual proteins as well as entire proteome samples. The utility of these methods is illustrated in examples that show the information that can be gained using these MS techniques.

磷酸化可以说是蛋白质中发生的最重要的翻译后修饰。磷酸化被用作信号来控制从基因表达到代谢的许多生理活动。识别蛋白质内的磷酸化位点历来是一项挑战，因为它需要放射性同位素标记或使用磷酸化特异性抗体。质谱 (MS) 的出现对定性和定量表征磷酸化蛋白质的能力产生了重大影响。在本文中，我们描述了表征单个蛋白质以及整个蛋白质组样品中磷酸化位点的 MS 方法。这些方法的效用在示例中进行了说明，这些示例显示了可以使用这些 MS 技术获得的信息。