Bioinformatics Analysis of Top-Down Mass Spectrometry Data with ProSight Lite.
Bioinformatics Analysis of Top-Down Mass Spectrometry Data with ProSight Lite.
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DOI：
10.1007/978-1-4939-6783-4_18
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Thomas PM
中科院分区：
文献类型：
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作者：
DeHart CJ;Fellers RT;Fornelli L;Kelleher NL;Thomas PM
传统的基于质谱的自下而上的蛋白质组学依赖于使用酶（通常是胰蛋白酶）来产生小肽（通常< 25个氨基酸长）。在自上而下的蛋白质组学中，蛋白质保持完整，并在质谱仪内直接测量。这种技术，虽然本质上比自下而上的蛋白质组学更简单，但产生的数据必须使用专门为这项工作设计的软件工具进行处理和分析。在本章中，我们将展示完整蛋白质光谱的分析，通过去卷积，deisotoping，和搜索与ProSight Lite，一个免费的，供应商不可知的工具，自上而下的质谱数据的分析。我们将说明两个完整的蛋白质片段化光谱的例子，并讨论迭代使用的软件来表征proteoforms和发现的翻译后修饰的网站。
Traditional bottom-up mass spectrometry-based proteomics relies on the use of an enzyme, often trypsin, to generate small peptides (typically < 25 amino acids long). In top-down proteomics, proteins remain intact and are directly measured within the mass spectrometer. This technique, while inherently simpler than bottom-up proteomics, generates data which must be processed and analyzed using software tools “purpose-built” for the job. In this chapter, we will show the analysis of intact protein spectra through deconvolution, deisotoping, and searching with ProSight Lite, a free, vendor-agnostic tool for the analysis of top-down mass spectrometry data. We will illustrate with two examples of intact protein fragmentation spectra and discuss the iterative use of the software to characterize proteoforms and discover the sites of post-translational modifications.