Quantitative analysis of complex protein mixtures using isotope-coded affinity tags
Quantitative analysis of complex protein mixtures using isotope-coded affinity tags
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DOI：
10.1038/13690
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发表时间：
1999-10-01
影响因子：
46.9
通讯作者：
Aebersold, R
中科院分区：
文献类型：
--
作者：
Gygi, SP;Rist, B;Aebersold, R
我们描述了一种对复杂混合物中的单个蛋白质进行准确定量和同时序列鉴定的方法。该方法基于一类称为同位素编码亲和标签 (ICAT) 的新型化学试剂和串联质谱法。使用这种策略，我们使用乙醇或半乳糖作为碳源，比较了酿酒酵母中的蛋白质表达。测量到的蛋白质表达差异与葡萄糖抑制条件下已知的酵母代谢功能相关。如果存在多个半胱氨酰残基，则该方法是多余的，并且相对定量非常准确，因为它基于稳定同位素稀释技术。 ICAT 方法应该提供一种广泛适用的方法来定量比较细胞和组织中的整体蛋白质表达。
We describe an approach for the accurate quantification and concurrent sequence identification of the individual proteins within complex mixtures. The method is based on a class of new chemical reagents termed isotope-coded affinity tags (ICATs) and tandem mass spectrometry. Using this strategy, we compared protein expression in the yeast Saccharomyces cerevisiae, using either ethanol or galactose as a carbon source. The measured differences in protein expression correlated with known yeast metabolic function under glucose-repressed conditions. The method is redundant if multiple cysteinyl residues are present, and the relative quantification is highly accurate because it is based on stable isotope dilution techniques. The ICAT approach should provide a widely applicable means to compare quantitatively global protein expression in cells and tissues.