MODIFICATION OF CYSTEINE RESIDUES FOR MASS SPECTROMETRY-BASED PROTEOMIC ANALYSIS: FACTS AND ARTIFACTS
MODIFICATION OF CYSTEINE RESIDUES FOR MASS SPECTROMETRY-BASED PROTEOMIC ANALYSIS: FACTS AND ARTIFACTS
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DOI：
10.18097/pbmc20206601018
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发表时间：
2020-01-01
影响因子：
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通讯作者：
Moshkovskii, S. A.
中科院分区：
文献类型：
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作者：
Kuznetsova, K. G.;Solovyeva, E. M.;Moshkovskii, S. A.
样品制备阶段的质谱学蛋白质组学分析包括人工还原半胱氨酸残基之间形成的蛋白质中的二硫键。这种键，当保存在它们的天然状态时，会使随后的酶水解和对研究结果的解释复杂化。为了防止二硫键的重新形成，半胱氨酸残基被特殊的基团保护，最常见的是烷基化。在这篇综述中，我们考虑了在样品制备过程中用来修饰半胱氨酸残基的方法，以及这一阶段可能出现的伪影。特别是，描述了烷化剂与其他氨基酸残基的不良反应。在蛋白质组分析中，用来保护半胱氨酸残基的最常见的烷基化化合物是碘乙酰胺。然而，对这一领域的文献分析表明，该试剂比其他使用的试剂，如氯乙酰胺和丙烯酰胺，会引起更多的不良反应。后者可推荐用于更广泛的用途。在综述中，我们还讨论了半胱氨酸残基修饰的特点及其对单核苷酸替代的翻译后修饰和蛋白质产物搜索效率的影响。
Mass spectrometric proteomic analysis at the sample preparation stage involves the artificial reduction of disulfide bonds in proteins formed between cysteine residues. Such bonds, when preserved in their native state, complicate subsequent enzymatic hydrolysis and interpretation of the research results. To prevent the re-formation of the disulfide bonds, cysteine residues are protected by special groups, most often by alkylation. In this review, we consider the methods used to modify cysteine residues during sample preparation, as well as possible artifacts of this stage. Particularly, adverse reactions of the alkylating agents with other amino acid residues are described. The most common alkylating compound used to protect cysteine residues in mass spectrometric proteomic analysis is iodoacetamide. However, an analysis of the literature in this area indicates that this reagent causes more adverse reactions than other agents used, such as chloroacetamide and acrylamide. The latter can be recommended for wider use. In the review we also discuss the features of the cysteine residue modifications and their influence on the efficiency of the search for post-translational modifications and protein products of single nucleotide substitutions.