Large-scale analysis of the yeast proteome by multidimensional protein identification technology
Large-scale analysis of the yeast proteome by multidimensional protein identification technology
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DOI：
10.1038/85686
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发表时间：
2001-03-01
影响因子：
46.9
通讯作者：
Yates, JR
中科院分区：
文献类型：
--
作者：
Washburn, MP;Wolters, D;Yates, JR
我们描述了一个很大程度上无偏的方法，快速和大规模的蛋白质组分析多维液相色谱，串联质谱，和数据库搜索的SEQUEST算法，命名为多维蛋白质鉴定技术（MudPIT）。将MudPIT应用于生长至对数中期的酿酒酵母菌株BJ 5460的蛋白质组，并产生了迄今为止最大的蛋白质组分析。共检测和鉴定了1,484种蛋白质。这些命中的分类证明了该技术检测和鉴定蛋白质组分析中很少见到的蛋白质的能力，包括低丰度蛋白质，如转录因子和蛋白激酶。此外，我们确定了131个蛋白质与三个或更多的预测跨膜结构域，这使我们能够映射的可溶性结构域的许多完整的膜蛋白。MudPIT可用于蛋白质组分析，并可专门应用于完整膜蛋白，以获得有关这类笨重蛋白质的详细生化信息。
We describe a largely unbiased method for rapid and large-scale proteome analysis by multidimensional liquid chromatography, tandem mass spectrometry, and database searching by the SEQUEST algorithm, named multidimensional protein identification technology (MudPIT). MudPIT was applied to the proteome of the Saccharomyces cerevisiae strain BJ5460 grown to mid-log phase and yielded the largest proteome analysis to date. A total of 1,484 proteins were detected and identified. Categorization of these hits demonstrated the ability of this technology to detect and identify proteins rarely seen in proteome analysis, including low-abundance proteins like transcription factors and protein kinases. Furthermore, we identified 131 proteins with three or more predicted transmembrane domains, which allowed us to map the soluble domains of many of the integral membrane proteins. MudPIT is useful for proteome analysis and may be specifically applied to integral membrane proteins to obtain detailed biochemical information on this unwieldy class of proteins.