Analysis of the mouse liver proteome using advanced mass spectrometry
Analysis of the mouse liver proteome using advanced mass spectrometry
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DOI：
10.1021/pr0605668
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发表时间：
2007-08-01
影响因子：
4.4
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Shi, Rong;Kumar, Chanchal;Mann, Matthias
我们报告了一个大规模的分析小鼠肝脏组织，包括一个新的分馏方法和高精度质谱技术。从20 mg冷冻组织中富集可溶性和膜蛋白的两个级分通过一维电泳分离，然后在混合线性离子阱（LTQ）-Orbitrap质谱仪上进行LC-MS/MS。2210蛋白质的可靠鉴定依赖于至少两种肽。我们将该蛋白质组与我们先前报道的细胞器图谱（Foster等Cell 2006，125，187-199）相结合，以产生3244种蛋白质的非常高置信度的小鼠肝脏蛋白质组。所鉴定的蛋白质代表肝脏蛋白质组，由于蛋白质的理化性质、亚细胞分布或生物学功能，没有可辨别的偏倚。47%的鉴定蛋白质被注释为膜结合的，35.3%，预测跨膜结构域。对于毒理学或临床研究中的潜在应用，我们证明了在单次运行中可以一致地识别1000多种蛋白质。
We report a large-scale analysis of mouse liver tissue comprising a novel fractionation approach and high-accuracy mass spectrometry techniques. Two fractions enriched for soluble and membrane proteins from 20 mg of frozen tissue were separated by one-dimensional electrophoresis followed by LC-MS/MS on the hybrid linear ion trap (LTQ)-Orbitrap mass spectrometer. Confident identification of 2210 proteins relied on at least two peptides. We combined this proteome with our previously reported organellar map (Foster et al. Cell 2006, 125, 187-199) to generate a very high confidence mouse liver proteome of 3244 proteins. The identified proteins represent the liver proteome with no discernible bias due to protein physicochemical properties, subcellular distribution, or biological function. Forty-seven percent of identified proteins were annotated as membrane-bound, and for 35.3%, transmembrane domains were predicted. For potential application in toxicology or clinical studies, we demonstrate that it is possible to consistently identify more than 1000 proteins in a single run.