串联质量标签用于比较和发现蛋白质组学。

Tandem Mass Tags for Comparative and Discovery Proteomics.

机构信息

Leibniz-Institut für Analytische Wissenschaften - ISAS - e.V., Bunsen-Kirchhoff-Straße 11, Dortmund, Germany.

Department of Chemistry, College of Physical Sciences, University of Aberdeen, Aberdeen, Scotland.

出版信息

Methods Mol Biol. 2021;2228:117-131. doi: 10.1007/978-1-0716-1024-4_9.

DOI:10.1007/978-1-0716-1024-4_9

PMID:33950487

Abstract

Relative or comparative proteomics provides valuable insights about the altered protein abundances across different biological samples in a single (labeled) or series (label-free) of LC-MS measurement(s). Chemical labeling of peptides using isobaric mass tags for identification and quantification of different proteomes simultaneously has become a routine in the so-called discovery proteomics in the past decade. One of the earliest isobaric tags-based technologies is TMT (tandem mass tags), which relies on the comparison of the unique "reporter ions" intensities for relative peptide/protein quantification. This differential labeling approach has evolved over time with respect to its multiplexing capability, i.e., from just 2 samples (TMTduplex) to 10 samples (TMT10plex) and a nowadays of up to 16 samples (TMTpro 16plex). Here, we describe a straightforward protocol to perform relatively deep proteome quantitative analyses using TMT10plex.

摘要

相对或比较蛋白质组学提供了有价值的见解，了解在单个（标记）或一系列（无标记） LC-MS 测量中不同生物样本中蛋白质丰度的变化。在过去十年中，使用等质量标记物对肽进行化学标记，以同时鉴定和定量不同的蛋白质组，已成为所谓的发现蛋白质组学中的常规方法。最早的基于等质量标记物的技术之一是 TMT（串联质量标签），它依赖于比较独特的“报告离子”强度来进行相对肽/蛋白质定量。这种差异标记方法在其多重检测能力方面不断发展，即从最初仅适用于 2 个样本（TMTduplex），发展到现在可用于 10 个样本（TMT10plex），甚至最高可达 16 个样本（TMTpro 16plex）。在这里，我们描述了一种使用 TMT10plex 进行相对深度蛋白质组定量分析的简单方案。