Oda Y, Huang K, Cross F R, Cowburn D, Chait B T
The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):6591-6. doi: 10.1073/pnas.96.12.6591.
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.
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，并确定单个蛋白质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的定量。该方法应用于检测野生型与突变型细胞群体中存在的蛋白质丰度差异，并鉴定与PAK相关的酵母Ste20蛋白激酶中体内磷酸化位点，这些位点特别依赖于G1细胞周期蛋白Cln2。本方法具有通用性，能在蛋白质表达和修饰水平上对细胞差异进行定量描述，从而提供对于理解复杂生物学现象至关重要的信息。
