Analysis of quantitative proteomic data generated via multidimensional protein identification technology
Analysis of quantitative proteomic data generated via multidimensional protein identification technology
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DOI：
10.1021/ac015704l
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发表时间：
2002-04-01
影响因子：
7.4
通讯作者：
Yates, JR
中科院分区：
文献类型：
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作者：
Washburn, MP;Ulaszek, R;Yates, JR
我们描述了通过多维蛋白质鉴定技术（MudPIT）对定量蛋白质组学样品的分析。将培养于N-14最小培养基或n -15富集最小培养基中的酿酒酵母菌株S288C的蛋白质组可溶性部分按比例混合并消化成复合肽混合物。采用一维反相色谱法和电喷雾电离飞行时间质谱法分析了1x N-14/lx N-15复合肽混合物，以证明在所使用的生长条件下N-15取代了N-14。将N-15掺入标记样品构象后，通过MudPIT分析由1 × N-14/1 × N-15复合多肽混合物、5 × N-14/1 × N-15复合多肽混合物和10 × 14N/ 1 × 15N复合多肽混合物组成的三个独立样品。我们通过分析在N-14或n -15富集的最小培养基中生长的酿酒酵母的可溶部分的1:1、5:1和10:1数据集来证明该系统的动态范围。所描述的方法提供了一种进行大规模定量蛋白质组学研究的准确方法。
We describe the analysis of quantitative proteomic samples via multidimensional protein identification technology (MudPIT. Ratio amounts of the soluble portion of the S. cerevisiae proteome from cultures of S. cerevisiae strain S288C grown in either N-14 minimal media or N-15-enriched minimal media were mixed and digested into a complex peptide mixture. A 1x N-14/lx N-15 complex peptide mixture was analyzed by single-dimensional reversed-phase chromatography and electrospray ionization quadrapole time-of-flight mass spectrometry in order to demonstrate the replacement of N-14 by N-15 under the growth conditions used. After conformation of the incorporation of N-15 into the labeled sample, three separate samples consisting of a 1 x N-14/ 1 x N-15 complex peptide mixture, a 5 x N-14/1 x N-15 complex peptide mixture, and a 10 x 14N/ 1 x 15N complex peptide mixture were analyzed via MudPIT. We demonstrate the dynamic range of the system by analyzing a 1: 1, 5: 1, and 10: 1 data set using the soluble portion from S. cerevisiae grown in either N-14 or N-15-enriched minimal media. The method described provides an accurate way to undertake a large-scale quantitative proteomic study.