Proteome analysis of low-abundance proteins using multidimensional chromatography and isotope-coded affinity tags
Proteome analysis of low-abundance proteins using multidimensional chromatography and isotope-coded affinity tags
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DOI：
10.1021/pr015509n
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发表时间：
2002-01-01
影响因子：
4.4
通讯作者：
Aebersold, R
中科院分区：
文献类型：
--
作者：
Gygi, SP;Rist, B;Aebersold, R
蛋白质组范围内的蛋白质鉴定和定量表达谱的有效性取决于常规获取低丰度蛋白质信息的分析方法的能力，因为这些通常具有重要的生物学意义。二维凝胶电泳，传统的蛋白质组分析方法，已被证明偏向于高表达的蛋白质。最近，引入了二维色谱法对未分离蛋白质样品消化产生的复杂肽混合物进行成分鉴定，并引入了同位素编码亲和标签（ICAT），以便通过质谱法对蛋白质混合物的成分进行准确定量。在这里，我们证明了同位素编码亲和蛋白标签和色氨酸混合物的多维色谱/质谱相结合能够检测和定量复杂样品中的低丰度蛋白质。
The effectiveness of proteome-wide protein identification and quantitative expression profiling is dependent on the ability of the analytical methodologies employed to routinely obtain information on low-abundance proteins, as these are frequently of great biological importance. Two-dimensional gel electrophoresis, the traditional method for proteome analysis, has proven to be biased toward highly expressed proteins. Recently, two-dimensional chromatography of the complex peptide mixtures generated by the digestion of unseparated protein samples has been introduced for the identification of their components, and isotope-coded affinity tags (ICAT) have been introduced to allow for accurate quantification of the components of protein mixtures by mass spectrometry. Here, we demonstrate that the combination of isotope coded affinity protein tags and multidimensional chromatography/mass spectrometry of tryptic peptide mixtures is capable of detecting and quantifying proteins of low abundance in complex samples.