Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, United States.
Integrated Biomedical Sciences Graduate Program, University of Notre Dame, Notre Dame, Indiana 46556, United States.
J Proteome Res. 2023 Feb 3;22(2):432-441. doi: 10.1021/acs.jproteome.2c00606. Epub 2023 Jan 18.
Bottom-up proteomics (BUP) produces rich data, but visualization and analysis are time-consuming and often require programming skills. Many tools analyze these data at the proteome-level, but fewer options exist for individual proteins. Sequence coverage maps are common, but do not proportion peptide intensity. Abundance-based visualization of sequence coverage facilitates detection of protein isoforms, domains, potential truncation sites, peptide "hot-spots", and localization of post-translational modifications (PTMs). Redundant stacked-sequence coverage is an important tool in designing hydrogen-deuterium exchange (HDX) experiments. Visualization tools often lack graphical and tabular-export of processed data which complicates publication of results. Quantitative peptide abundance across amino acid sequences is an essential and missing tool in proteomics toolkits. Here we created PrIntMap-R, an online application that only requires peptide files from a database search and FASTA protein sequences. PrIntMap-R produces a variety of plots for quantitative visualization of coverage; annotation of specific sequences, PTM's, and comparisons of one or many samples overlaid with calculated fold-change or several intensity metrics. We show use-cases including protein phosphorylation, identification of glycosylation, and the optimization of digestion conditions for HDX experiments. PrIntMap-R is freely available, open source, and can run online with no installation, or locally by downloading source code from GitHub.
自下而上蛋白质组学(BUP)产生丰富的数据,但可视化和分析既耗时又常常需要编程技能。许多工具可在蛋白质组水平上分析这些数据,但针对个别蛋白质的选择较少。序列覆盖图很常见,但不能按比例显示肽强度。基于丰度的序列覆盖可视化有助于检测蛋白质同工型、结构域、潜在截断位点、肽“热点”以及翻译后修饰(PTM)的定位。冗余堆叠序列覆盖是设计氢氘交换(HDX)实验的重要工具。可视化工具通常缺乏处理后数据的图形和表格导出,这使得结果的发表变得复杂。定量肽丰度在整个氨基酸序列中是蛋白质组学工具包中必不可少但缺失的工具。在这里,我们创建了 PrIntMap-R,这是一个在线应用程序,仅需要数据库搜索和 FASTA 蛋白质序列的肽文件。PrIntMap-R 生成各种覆盖范围的定量可视化图;注释特定序列、PTM 以及比较一个或多个样本,叠加计算的倍数变化或几个强度指标。我们展示了一些用例,包括蛋白质磷酸化、糖基化鉴定以及优化 HDX 实验的消化条件。PrIntMap-R 是免费的、开源的,可以在线运行,无需安装,也可以从 GitHub 下载源代码在本地运行。