The role of sulfur and sulfur isotope dilution analysis in quantitative protein analysis
The role of sulfur and sulfur isotope dilution analysis in quantitative protein analysis
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DOI：
10.1007/s00216-007-1607-2
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发表时间：
2008
影响因子：
4.3
通讯作者：
C. Rappel;D. Schaumlöffel
中科院分区：
文献类型：
--
作者：
C. Rappel;D. Schaumlöffel
硫元素在所有天然蛋白质组中几乎无所不在，在蛋白质定量中起着关键作用。它与氨基酸半胱氨酸和蛋氨酸结合，在经典的基于电喷雾或MALDI质谱的定量蛋白质组学方法中被用作许多蛋白质标记反应的靶标。本文讨论了硫同位素稀释分析（IDA）电感耦合等离子体质谱（ICP-MS）用于绝对蛋白质定量的潜力和局限性。由于近年来ICP-MS测量硫同位素比的灵敏度和准确性的提高，使这种方法的发展成为可能。ICP-MS的独特之处是化合物独立电离，使化合物（种）非特异性硫IDA成为可能。这样做的主要优点是，只要样品中的每个肽或蛋白质在色谱或电泳中完全分离并且它们的特性已知，就只需要一个通用硫标准（即一个同位素标记的硫峰）来量化样品中的每个肽或蛋白质。这种方法的原理是从文献中选择的例子说明。讨论了P/S和金属/S比值测定蛋白磷酸化程度和金属蛋白化学计量学的相关领域。讨论了新出现的新领域和未来趋势，如金属标签的蛋白质衍生化，以提高ICP-MS中蛋白质检测的灵敏度。图硫在蛋白质定量中的关键作用
The element sulfur is almost omnipresent in all natural proteomes and plays a key role in protein quantification. Incorporated in the amino acids cysteine and methionine, it has been served as target for many protein-labeling reactions in classic quantitative proteomic approaches based on electrospray or MALDI mass spectrometry. This critical review discusses the potential and limitations of sulfur isotope dilution analysis (IDA) by inductively coupled plasma—mass spectrometry (ICP-MS) for absolute protein quantification. The development of this approach was made possible due to the improved sensitivity and accuracy of sulfur isotope ratio measurement by ICP-MS in recent years. The unique feature of ICP-MS, compound-independent ionization, enables compound (species)-unspecific sulfur IDA. This has the main advantage that only one generic sulfur standard (i.e., one isotopically labeled sulfur spike) is required to quantify each peptide or protein in a sample provided that they are completely separated in chromatography or electrophoresis and that their identities are known. The principles of this approach are illustrated with selected examples from the literature. The discussion includes also related fields of P/S and metal/S ratio measurements for the determination of phosphorylation degrees of proteins and stoichiometries in metalloproteins, respectively. Emerging new areas and future trends such as protein derivatization with metal tags for improved sensitivity of protein detection in ICP-MS are discussed.FigureThe key role of sulfur in protein quantification