Zybailov Boris, Mosley Amber L, Sardiu Mihaela E, Coleman Michael K, Florens Laurence, Washburn Michael P
Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110, USA.
J Proteome Res. 2006 Sep;5(9):2339-47. doi: 10.1021/pr060161n.
We have devised an approach for analyzing shotgun proteomics datasets based on the normalized spectral abundance factor that can be used for quantitative proteomics analysis. Three biological replicates of samples enriched for plasma membranes were isolated from S. cerevisiae grown in 14N-rich media and 15N-minimal media and analyzed via quantitative multidimensional protein identification technology. The natural log transformation of NSAF values from S. cerevisiae cells grown in 14N YPD media and 15N-minimal media had a normal distribution. The t-test analysis demonstrated 221 of 1316 proteins were significantly overexpressed in one or the other growth conditions with a p value <0.05. Notably, amino acid transporters were among the 14 membrane proteins that were significantly upregulated in cells grown in minimal media, and we functionally validated these increases in protein expression with radioisotope uptake assays for selected proteins.
我们设计了一种基于标准化谱丰度因子分析鸟枪法蛋白质组学数据集的方法，该方法可用于定量蛋白质组学分析。从在富含14N的培养基和15N基本培养基中生长的酿酒酵母中分离出三份富含质膜的样品生物学重复，并通过定量多维蛋白质鉴定技术进行分析。在14N YPD培养基和15N基本培养基中生长的酿酒酵母细胞的NSAF值的自然对数转换呈正态分布。t检验分析表明，1316种蛋白质中有221种在一种或另一种生长条件下显著过表达，p值<0.05。值得注意的是，氨基酸转运蛋白是在基本培养基中生长的细胞中显著上调的14种膜蛋白之一，我们通过对选定蛋白质的放射性同位素摄取试验在功能上验证了这些蛋白质表达的增加。
