Mix-and-Match Proteomics: Using Advanced Iodoacetyl Tandem Mass Tag Multiplexing To Investigate Cysteine Oxidation Changes with Respect to Protein Expression.
Mix-and-Match Proteomics: Using Advanced Iodoacetyl Tandem Mass Tag Multiplexing To Investigate Cysteine Oxidation Changes with Respect to Protein Expression.
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混合匹配蛋白质组学：使用先进的碘乙酰串联质量标签多重技术研究半胱氨酸氧化与蛋白质表达的关系。
DOI：
10.1021/acs.analchem.8b02517
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发表时间：
2018
影响因子：
7.4
通讯作者：
Prakash AS
中科院分区：
文献类型：
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作者：
Prakash AS
半胱氨酸氧化还原状态被认为是影响蛋白质结构和/或功能的关键生物学因素之一。蛋白质氧化还原状态的改变已被证明会引起显著的生理变化，并会使蛋白质对氧化应激的敏感性发生变化。蛋白质氧化还原状态的变化被认为是疾病、细胞异常和环境变化的重要中介，因此它们的特征是有意义的。同位素或等压标记，然后样品多路传输和分析的液体色谱串联质谱(LC-MS/MS)允许在几个样品之间的蛋白质表达水平或蛋白质氧化还原状态的相对比较。将蛋白质表达水平和氧化还原状态的分析结合到一个分析中将增加额外的维度，并允许蛋白质氧化还原随蛋白质丰度的变化标准化。为了实现这一点，我们开发了一种定量工作流程，使用商业上可用的半胱氨酸-反应串联质量标签(IodoTMT)来差异标记半胱氨酸残基，并将其应用于两个先前对氧化应激表现出不同反应的利什曼原虫。将单独标记的样本以不同的组合组合在一起，创建多个六链样本，以研究半胱氨酸氧化与整体蛋白质表达之间的关系，并提供有关每个细胞系中蛋白质氧化水平的信息。结果突出了在两种细胞系之间差异表达和/或具有显著氧化还原变化的11种蛋白质。这种先进的多路复用方法有效地展示了串联质量标签的灵活性，以及如何使用它们来最大限度地获取信息量。
Cysteine redox state has been identified as one of the key biological influences behind protein structure and/or function. Altered protein redox state has been shown to cause significant physiological changes and can leave proteins with changed sensitivity to oxidative stress. Protein redox-state changes are recognized as an important mediator of disease, cellular abnormalities, and environmental changes, and therefore their characterization is of interest. Isotopic or isobaric labeling followed by sample multiplexing and analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS) allows relative comparison of protein expression levels or of protein redox states between several samples. Combining analysis of protein expression level and redox state into one analysis would add an extra dimension and permit the normalization of protein redox changes with protein abundance. To achieve this, we have developed a quantitation workflow that uses commercially available cysteine-reactive tandem mass tags (iodoTMT) to differentially label cysteine residues, and we have applied it to twoLeishmania mexicanacell lines that have previously shown different responses to oxidative stress. The individually labeled samples have been pooled in different combinations to create multiple sixplex samples in order to study the relationship between cysteine oxidation and overall protein expression, as well as providing information about protein oxidation levels in each cell line. The results highlight 11 proteins that are differentially expressed between the two cell lines and/or have significant redox changes. This advanced multiplexing method effectively demonstrates the flexibility of tandem mass tags and how they can be used to maximize the amount of information that can be acquired.
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影响因子：
3.8
作者：
Daneshvar H;Namazi MJ;Kamiabi H;Burchmore R;Cleaveland S;Phillips S
通讯作者：
Phillips S
影响因子：
3.3
作者：
Daneshvar, Hamid;Wyllie, Susan;Burchmore, Richard
通讯作者：
Burchmore, Richard
影响因子：
6.4
作者：
Daneshvar, H;Coombs, GH;Phillips, RS
通讯作者：
Phillips, RS
0kajima, T 等人：“小鼠 GD la/Gtlaa/Gqlba 合酶 (ST6GalNAcVH1) 基因的分子克隆和表达。”J.
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发表时间：
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影响因子：
--
作者：
通讯作者：
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影响因子：
4
作者：
Romao, Susana;Castro, Helena;Tomas, Ana M.
通讯作者：
Tomas, Ana M.