Taking the Shortcut for High-Throughput Shotgun Proteomic Analysis of Bacteria
Taking the Shortcut for High-Throughput Shotgun Proteomic Analysis of Bacteria
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DOI：
10.1007/978-1-4939-1261-2_16
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Armengaud, Jean
中科院分区：
文献类型：
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作者：
Hartmann, Erica Marie;Allain, Francois;Armengaud, Jean
目前，蛋白质组学工具能够建立样品中最丰富蛋白质的完整列表，为高分辨率研究任何细菌的生理提供了机会，这些细菌的基因组序列是可用的。对于一个全面的列表，蛋白质应该首先被分解成部分，然后被胰蛋白酶水解。所得到的肽混合物由高通量串联质谱仪分析，该质谱仪记录每个部分的数千个MS/MS光谱。然后将这些光谱分配给肽，作为蛋白质存在的证据。除了生成蛋白质鉴定列表外，这种蛋白质组学的捷径还使用每种蛋白质记录的光谱数量来量化观察结果。在这里，我们描述了一种最简单的细菌高通量蛋白质组学的样品制备方法，以及随后的数据处理，以提取基于光谱计数方法的定量信息。
Currently, proteomic tools are able to establish a complete list of the most abundant proteins present in a sample, providing the opportunity to study at high resolution the physiology of any bacteria for which the genome sequence is available. For a comprehensive list, proteins should be first resolved into fractions that are then proteolyzed by trypsin. The resulting peptide mixtures are analyzed by a high-throughput tandem mass spectrometer that records thousands of MS/MS spectra for each fraction. These spectra are then assigned to peptides, which are used as evidence of the existence of proteins. In addition to generating a list of protein identifications, this shortcut to proteomics uses the number of spectra recorded for each protein to quantify the observations. Here, we describe one of the most simple sample preparation methods for high-throughput proteomics of bacteria, as well as the subsequent data processing to extract quantitative information based on the spectral count approach.