Department of Chemistry, Università degli Studi di Roma La Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy.
Department of Chemistry, Università degli Studi di Roma La Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy.
Talanta. 2020 Nov 1;219:121262. doi: 10.1016/j.talanta.2020.121262. Epub 2020 Jun 16.
Short peptides are of extreme interest in clinical and food research fields, nevertheless they still represent a crucial analytical issue. The main aim of this paper was the development of an analytical platform for a considerable advancement in short peptides identification. For the first time, short sequences presenting both natural and post-translationally modified amino acids were comprehensively studied thanks to the generation of specific databases. Short peptide databases had a dual purpose. First, they were employed as inclusion lists for a suspect screening mass-spectrometric analysis, overcoming the limits of data dependent acquisition mode and allowing the fragmentation of such low-abundance substances. Moreover, the databases were implemented in Compound Discoverer 3.0, a software dedicated to the analysis of short molecules, for the creation of a data processing workflow specifically dedicated to short peptide tentative identification. For this purpose, a detailed study of short peptide fragmentation pathways was carried out for the first time. The proposed method was applied to the study of short peptide sequences in enriched urine samples and led to the tentative identification more than 200 short natural and modified short peptides, the highest number ever reported.
短肽在临床和食品研究领域极具研究价值，但它们仍然是一个关键的分析问题。本文的主要目的是开发一个分析平台，以实现短肽鉴定的重大进展。这是首次全面研究既包含天然氨基酸又包含翻译后修饰氨基酸的短序列，这要归功于特定数据库的生成。短肽数据库有两个目的。首先，它们被用作候选筛选质谱分析的包含列表，克服了数据依赖采集模式的限制，并允许对这些低丰度物质进行碎片化。此外，数据库被集成到 Compound Discoverer 3.0 中，这是一款专门用于分析短分子的软件，用于创建专门用于短肽初步鉴定的数据处理工作流程。为此，首次对短肽的碎片化途径进行了详细研究。该方法被应用于富含尿液样本中的短肽序列研究，首次鉴定出 200 多种天然和修饰的短肽，这是迄今为止报道的最高数量。
