N-t-butyliodoacetamide and iodoacetanilide:: two new cysteine alkylating reagents for relative quantitation of proteins
N-t-butyliodoacetamide and iodoacetanilide:: two new cysteine alkylating reagents for relative quantitation of proteins
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DOI：
10.1002/rcm.1286
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发表时间：
2004-01-01
影响因子：
2
通讯作者：
Corthals, GL
中科院分区：
文献类型：
--
作者：
Pasquarello, C;Sanchez, JC;Corthals, GL
本文介绍了两种新的烷基化试剂N-叔丁基-2-碘乙酰胺（N-叔丁基碘乙酰胺）和2-碘-N-苯基乙酰胺（碘乙酰苯胺）的合成及应用。合成N-叔丁基碘乙酰胺和碘乙酰苯胺，使其纯度达到d（0）-轻形式和d（9）-和d（5）-重形式。新合成的试剂通过烷基化反应与含有半胱氨酸的肽共价结合。5和9 Da的质量差避免了同位素分布重叠的可能问题。对于每个烷基化的半胱氨酸，对于d（0）-轻形式的N-叔丁基碘乙酰胺和碘乙酰苯胺，肽质量分别增加113和133 Da的倍数。因此，这些试剂可以取代现有基于蛋白质组学的应用中的常见烷基化试剂。烷基化肽的质量在与氨基酸相同的质量范围内增加，并且仍然适用于串联质谱（MS/MS）数据采集和分析。这些化合物使用简单，衍生化是基于广泛应用的烷基化程序。初步结果表明，这些试剂可以应用于蛋白质定量和鉴定肽质量指纹图谱和/或MS/MS技术。使用这些化学品和建议的工作流程可以对整个蛋白质样品进行定量分析，并实现对可能含有潜在翻译后修饰的肽的访问。在MS之前结合基质辅助激光解吸/电离（MALDI）接口的其他方法可以利用这些化学品，例如分子扫描仪。版权所有（C）2003约翰威利父子有限公司。
The synthesis and application of two new alkylating reagents, N-tert-butyl-2-iodoacetamide (N-t-butyliodoacetamide) and 2-iodo-N-phenylacetamide (iodoacetanilide), are described. N-t-Butyliodoacetamide and iodoacetanilide were synthesised to purity in their d(0)-light and in their respective d(9)- and d(5)-heavy forms. The newly synthesised reagents are covalently bound to peptides containing cysteines via an alkylation reaction. The mass differences of 5 and 9 Da avoid possible problems of overlapping isotope distribution. For each alkylated cysteine a peptide mass increases, respectively, by a multiple of 113 and 133 Da for the d(0)-light form of N-t-butyliodoacetamide and iodoacetanilide. These reagents can therefore replace common alkylating reagents in existing proteomics-based applications. Alkylated peptides increase in mass in the same mass range as amino acids and remain suitable for tandem mass spectrometry (MS/MS) data acquisition and analysis. The compounds are simple to use and derivatisation is based on widely applied alkylating procedures. Preliminary results show that these reagents can be applied for both protein quantitation and identification by peptide mass finger printing and/or MS/MS techniques. Using these chemicals and the suggested workflow enables the quantitative analysis of the whole protein sample and realises access to peptides that may contain potential post-translational modifications. Other approaches that incorporate a matrix-assisted laser desorption/ionisation (MALDI) interface prior to MS can take advantage of these chemicals, such as the molecular scanner. Copyright (C) 2003 John Wiley Sons, Ltd.