构建用于鉴定蓖麻毒素的串联质谱库。,Journal of Proteome Research

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构建用于鉴定蓖麻毒素的串联质谱库。
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-10-14 , DOI: 10.1021/acs.jproteome.9b00377
Isabelle O'Bryon ₁ , Abigail E Tucker ₁ , Brooke L D Kaiser ₁ , Karen L Wahl ₁ , Eric D Merkley ₁

Affiliation

蓖麻毒素是一种在蓖麻籽中发现的蛋白质，是一种致命毒素，被指定为2类选择剂，蓖麻毒蛋白中毒的案例相对普遍。许多检测蛋白质毒素的方法（例如蓖麻毒素）使用靶向液相色谱-串联质谱（LC-MS / MS）来鉴定毒素肽，通常是胰蛋白酶肽。也已经成功地使用了非针对性的方法。但是，在法医学实践中，使用非目标蛋白质组学方法（包括数据库搜索）进行肽和蛋白质鉴定并不常见，对于法医学工作者来说可能并不熟悉。在这里，我们提出了一种创建胰蛋白酶蓖麻毒素肽谱库的方法，并通过谱库搜索将这些库用于蓖麻毒素鉴定，由于光谱库在小分子鉴定中的使用，对于法医科学家可能更熟悉。肽谱库可直接与真实标准（法医分析的关键要素）进行比较，但以前从未在法医环境中使用过。为了构建这些光谱库，使用胰蛋白酶消化了两个纯蓖麻毒蛋白样品（一个来自建议的标准参考物质），并使用标准shot弹枪LC-MS / MS协议进行了分析。根据从四个数据库搜索工具的过滤搜索结果中识别出的胰蛋白酶肽段创建谱库。然后将该库用于使用SpectraST进行法医学鉴定的蓖麻籽提取物的搜索。这些蓖麻籽样品是使用现实蓖麻毒素病例中常见的粗制方法制得的。分析表明，与MS-GF +和Sequest plus Percolator数据库搜索相比，谱库搜索可从粗蓖麻籽样品中鉴定出更多肽。这些结果是使用光谱库搜索来首次检测法医相关样品中蛋白质毒素的结果，表明推定的蓖麻毒素肽光谱与库谱的计算比较可能是检测未知样品中蓖麻毒素的有效方法。数据可通过ProteomeXchange获得，其标识符为PXD013711。这表明推定的蓖麻毒素肽谱与库谱的计算比较可能是检测未知样品中蓖麻毒素的有效方法。数据可通过ProteomeXchange获得，其标识符为PXD013711。这表明推定的蓖麻毒素肽谱与库谱的计算比较可能是检测未知样品中蓖麻毒素的有效方法。数据可通过ProteomeXchange获得，其标识符为PXD013711。

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Constructing a Tandem Mass Spectral Library for Forensic Ricin Identification.

Ricin, a protein found in castor seeds, is a lethal toxin that is designated as a category 2 select agent, and cases of attempted ricin poisoning are relatively common. Many methods to detect protein toxins such as ricin use targeted liquid chromatography–tandem mass spectrometry (LC–MS/MS) to identify toxin peptides, usually tryptic peptides. The successful use of untargeted methods has also been reported. However, the use of untargeted proteomics methods, including database search, for peptide and protein identification is less common in forensic practice and may be unfamiliar to forensic science practitioners. Here, we propose a method to create spectral libraries of tryptic ricin peptides and use these libraries for ricin identification by spectral library search, which may be more familiar to forensic scientists because of the use of spectral libraries in small molecule identification. Peptide spectral libraries offer a direct comparison to an authentic standard, a key element of forensic analysis, but have not previously been used in a forensic context. To construct these spectral libraries, two pure ricin samples (one from a proposed standard reference material) were digested with trypsin and analyzed using a standard shotgun LC–MS/MS protocol. Spectral libraries were created from resulting tryptic peptides identified from filtered search results from four database search tools. The library was then used in a search using SpectraST on forensically realistic castor seed extracts. These castor seed samples were made using the crude methods commonly encountered in real-world ricin cases. Analysis showed that the spectral library search resulted in more peptides identified from crude castor seed samples compared to MS-GF+ and Sequest plus Percolator database searches. These results, the first published use of spectral library search to detect protein toxins in forensically relevant samples, suggest that computational comparison of putative ricin peptide spectra to library spectra can be an effective method to detect ricin in an unknown sample. Data are available via ProteomeXchange with identifier PXD013711.

更新日期：2019-10-14

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