Accurate quantitation of protein expression and site-specific phosphorylation
Accurate quantitation of protein expression and site-specific phosphorylation
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DOI：
10.1073/pnas.96.12.6591
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发表时间：
1999-06-08
影响因子：
11.1
通讯作者：
Chait, BT
中科院分区：
文献类型：
--
作者：
Oda, Y;Huang, K;Chait, BT
描述了一种基于质谱的方法，用于同时鉴定和定量单个蛋白质，并用于确定单个蛋白质上特定位点修饰水平的变化。通过使用全细胞稳定同位素标记实现准确定量。该方法用于检测野生型与突变型细胞群中存在的蛋白质丰度差异，并用于鉴定pak相关酵母Ste20蛋白激酶中特异性依赖于G1细胞周期蛋白Cln2的体内磷酸化位点。目前的方法是通用的，并在蛋白质表达和修饰水平上提供了细胞差异的定量描述。从而提供对理解复杂生物现象至关重要的信息。
A mass spectrometry-based method is described for simultaneous identification and quantitation of individual proteins and for determining changes in the levels of modifications at specific sites on individual proteins. Accurate quantitation is achieved through the use of whole-cell stable isotope labeling. This approach was applied to the detection of abundance differences of proteins present in wild-type versus mutant cell populations and to the identification of in vivo phosphorylation sites in the PAK-related yeast Ste20 protein kinase that depend specifically on the G1 cyclin Cln2, The present method is general and affords a quantitative description of cellular differences at the level of protein expression and modification, thus providing information that is critical to the understanding of complex biological phenomena.