Gygi S P, Rist B, Gerber S A, Turecek F, Gelb M H, Aebersold R
Department of Molecular Biotechnology, University of Washington, Box 357730, Seattle WA 98195-7730, USA.
Nat Biotechnol. 1999 Oct;17(10):994-9. doi: 10.1038/13690.
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.
我们描述了一种用于准确量化和同时鉴定复杂混合物中单个蛋白质序列的方法。该方法基于一类称为同位素编码亲和标签（ICATs）的新型化学试剂和串联质谱法。使用这种策略，我们比较了以乙醇或半乳糖作为碳源的酿酒酵母中的蛋白质表达。所测得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酸残基，该方法会出现冗余情况，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方法应该能提供一种广泛适用的手段，用于定量比较细胞和组织中的整体蛋白质表达。
