Proteome-wide cellular protein concentrations of the human pathogen Leptospira interrogans.
Proteome-wide cellular protein concentrations of the human pathogen Leptospira interrogans.
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DOI：
10.1038/nature08184
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发表时间：
2009-08-06
期刊：
影响因子：
64.8
通讯作者：
Aebersold R
中科院分区：
文献类型：
--
作者：
Malmström J;Beck M;Schmidt A;Lange V;Deutsch EW;Aebersold R
基于质谱的相对蛋白质组定量方法对生命科学研究产生了广泛的影响。然而，重要的研究方向，特别是那些涉及生物过程的数学建模和模拟，也严重依赖于绝对的定量数据，即表达的蛋白质浓度作为细胞状态的函数的知识。到目前为止，蛋白质组的显著部分（73%）的绝对蛋白质浓度测量仅来自遗传改变的S。酿酒酵母细胞，这是一种不能直接从酵母移植到其他物种的技术。在这项研究中，我们开发并应用了一种基于质谱的策略来确定绝对量，即细胞群中每个细胞的蛋白质拷贝数的平均数，对于遗传上未受干扰的细胞中的蛋白质组的显著部分。将该技术应用于人类病原体问号钩端螺旋体（一种负责钩端螺旋体病的螺旋体），我们从不同状态的细胞中产生了83%的质谱可检测蛋白质组的绝对蛋白质丰度尺度。利用L.为了验证问号线虫，我们使用冷冻电子断层扫描（cryoET）形态学测量来在单细胞水平验证通过质谱法在细胞群体上确定的所选蛋白质的平均绝对丰度值。由于该策略相对快速且适用于任何细胞类型，我们预计它将成为定量生物学和系统生物学的基石。
Mass spectrometry based methods for relative proteome quantification have broadly impacted life science research. However, important research directions, particularly those involving mathematical modeling and simulation of biological processes, also critically depend on absolutely quantitative data, i.e. knowledge of the concentration of the expressed proteins as a function of cellular state. Until now, absolute protein concentration measurements of a significant fraction of the proteome (73%) have only been derived from genetically altered S. cerevisiae cells, a technique that is not directly portable from yeast to other species. In this study we developed and applied a mass spectrometry based strategy to determine the absolute quantity i.e. the average number of protein copies per cell in a cell population, for a significant fraction of the proteome in genetically unperturbed cells. Applying the technology to the human pathogen Leptospira interrogans, a spirochete responsible for Leptospirosis, we generated an absolute protein abundance scale for 83% of the mass spectrometry detectable proteome, from cells at different states. Taking advantage of the unique cellular dimensions of L. interrogans, we used cryo electron tomography (cryoET) morphological measurements to verify at the single cell level the average absolute abundance values of selected proteins determined by mass spectrometry on a population of cells. As the strategy is relatively fast and applicable to any cell type we expect that it will become a cornerstone of quantitative biology and systems biology.
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