Detection of Unknown Chemical Adduct Modifications on Proteins: From Wet to Dry Laboratory
Detection of Unknown Chemical Adduct Modifications on Proteins: From Wet to Dry Laboratory
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DOI：
10.1007/978-1-4939-9232-4_8
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发表时间：
2019-01-01
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影响因子：
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通讯作者：
Acosta-Martin, Adelina E.
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文献类型：
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作者：
Antinori, Paola;Michelot, Theo;Acosta-Martin, Adelina E.
蛋白质上化学加合物的检测和表征越来越受到人们的关注。在这里，我们描述了一种循序渐进的程序，以确定未知的化学加合物修饰蛋白质与给定的活性化合物相互作用的结果。该方案可分为两个同样重要的部分：(1)湿实验室工作，以产生高质量的体外修饰蛋白质的质谱学(MS)数据；(2)干实验室工作，分析所产生的MS数据，并对加合物的化学组成和氨基酸定位提供高度可信的定性和定量结果。该方法适用于在LC-MS/MS实验中可检测到的任何形成共价蛋白质加合物的药物或化学化合物的研究。
The detection and characterization of chemical adducts on proteins is of increasing interest. Here, we described a step-by-step procedure to identify unknown chemical adduct modifications on proteins resulting from the interaction with a given reactive compound. The protocol can be divided into two equally important parts: (1) the wet laboratory work, to produce high quality mass spectrometry (MS) data of in vitro modified proteins and (2) the dry laboratory work, to analyze the generated MS data and provide highly confident qualitative and quantitative results on the chemical composition and amino acid localization of adducts. This protocol is applicable to the study of any pharmaceutical or chemical compound forming covalent protein adducts, detectable in LC-MS/MS experiments.