Tan Feng, Zhang Yangjun, Wang Jinglan, Wei Junying, Cai Yun, Qian Xiaohong
State Key Laboratory of Proteomics-Beijing Proteome Research Center-Beijing Institute of Radiation Medicine, Beijing, China.
J Mass Spectrom. 2008 May;43(5):628-32. doi: 10.1002/jms.1362.
In this article, an effective method for dephosphorylation of phosphopeptides by cerium oxide is described. The dephosphorylation activity of cerium oxide was evaluated by two standard phosphopeptides and the phosphopeptides in digests of phosphoprotein alpha-casein and beta-casein. Results showed that the dephosphorylation of all the phosphopeptides was completed in 10 min, and temperature had little effect on the dephosphorylation, the dephosphorylation could be carried out at 0 degrees C, room temperature and 37 degrees C. The dephosphorylation mediated by cerium oxide can be attributed to Lewis acid and nucleophile activations. Advantages of using cerium oxide as catalyst for the dephosphorylation include: safe, simple, high catalytic activity, and no precise control of the treatment temperature. The method is valid for the phosphorylation of Ser, Thr and Tyr, and can be used for phosphoprotein analysis.
本文描述了一种通过氧化铈对磷酸肽进行去磷酸化的有效方法。通过两种标准磷酸肽以及磷蛋白α-酪蛋白和β-酪蛋白消化物中的磷酸肽来评估氧化铈的去磷酸化活性。结果表明,所有磷酸肽的去磷酸化在10分钟内完成,温度对去磷酸化影响很小,去磷酸化可在0℃、室温及37℃下进行。氧化铈介导的去磷酸化可归因于路易斯酸和亲核试剂的活化作用。使用氧化铈作为去磷酸化催化剂的优点包括:安全、简单、催化活性高,且无需精确控制处理温度。该方法对丝氨酸、苏氨酸和酪氨酸的磷酸化有效,可用于磷蛋白分析。