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Proteome-wide cellular protein concentrations of the human pathogen Leptospira interrogans

机译：人类病原体钩端螺旋体的蛋白质组蛋白全细胞蛋白质浓度

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摘要

Mass-spectrometry-based methods for relative proteome quantification have broadly affected life science research. However, important research directions, particularly those involving mathematical modelling and simulation of biological processes, also critically depend on absolutely quantitative data-that is, knowledge of the concentration of the expressed proteins as a function of cellular state. Until now, absolute protein concentration measurements of a considerable fraction of the proteome (73%) have only been derived from genetically altered Saccharontyces cerevisiae cells, a technique that is not directly portable from yeast to other species. Here we present a mass-spectrometry-based strategy to determine the absolute quantity, that is, the average number of protein copies per cell in a cell population, for a large 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 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. Because 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.
机译：基于质谱的相对蛋白质组定量方法已广泛影响生命科学研究。但是，重要的研究方向，尤其是那些涉及数学建模和生物过程模拟的研究方向，也严格地依赖于绝对定量的数据，即关于表达的蛋白质浓度随细胞状态变化的知识。到目前为止，蛋白质组中相当一部分蛋白质（73％）的绝对蛋白质浓度测量仅来自基因改变的酿酒酵母细胞，该技术不能直接从酵母移植到其他物种。在这里，我们提出了一种基于质谱的策略，用于确定绝对数量，即细胞群体中每个细胞的蛋白质拷贝的平均数量，以分析遗传上不受干扰的细胞中蛋白质组的大部分。将该技术应用于人类病原体钩端螺旋体螺线虫，我们从不同状态的细胞中产生了可检测质谱的蛋白质组的83％的绝对蛋白质丰度标度。利用问号乳酸菌独特的细胞大小，我们使用了冷冻电子断层扫描形态学测量方法，以在单细胞水平上验证通过质谱法在细胞群上确定的所选蛋白质的平均绝对丰度值。由于该策略相对较快并且适用于任何细胞类型，因此我们希望它将成为定量生物学和系统生物学的基石。

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  • 来源
    《Nature》 |2009年第7256期|762-765|共4页
  • 作者

    Johan Malmstroem; Martin Beck; Alexander Schmidt; Vinzenz Lange; Eric W. Deutsch; Ruedi Aebersold;

  • 作者单位

    Institute of Molecular Systems Biology, ETH Zurich (Swiss Federal Institute of Technology), Wolfgang Pauli-Strasse 16, CH-8093 Zurich, Switzerland;

    Institute of Molecular Systems Biology, ETH Zurich (Swiss Federal Institute of Technology), Wolfgang Pauli-Strasse 16, CH-8093 Zurich, Switzerland;

    Institute of Molecular Systems Biology, ETH Zurich (Swiss Federal Institute of Technology), Wolfgang Pauli-Strasse 16, CH-8093 Zurich, Switzerland Competence Center for Systems Physiology and Metabolic Diseases, CH-8093 Zurich, Switzerland;

    Institute of Molecular Systems Biology, ETH Zurich (Swiss Federal Institute of Technology), Wolfgang Pauli-Strasse 16, CH-8093 Zurich, Switzerland Competence Center for Systems Physiology and Metabolic Diseases, CH-8093 Zurich, Switzerland;

    Institute for Systems Biology, 1441 North 34th Street, Seattle, Washington 98103-8904, USA;

    Institute of Molecular Systems Biology, ETH Zurich (Swiss Federal Institute of Technology), Wolfgang Pauli-Strasse 16, CH-8093 Zurich, Switzerland Competence Center for Systems Physiology and Metabolic Diseases, CH-8093 Zurich, Switzerland Institute for Systems Biology, 1441 North 34th Street, Seattle, Washington 98103-8904, USA Faculty of Science, University of Zurich, CH-8057 Zurich, Switzerland;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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