An accurate mass tag strategy for quantitative and high-throughput proteome measurements.

摘要: We describe and demonstrate a global strategy that extends the sensitivity, dynamic range, comprehensiveness throughput of proteomic measurements based upon use polypeptide''accurate mass tags'' (AMTs) produced by protein enzymatic digestion. The two stage exploits Fourier transform ion cyclotron resonance spectrometry (FTICR) to first validate polypeptide AMTs for specific organism, tissue or cell type from''potential identified using conventional tandem (MS/MS) methods, providing basis subsequent without need MS/MS. A single high resolution capillary liquid chromatography separation combined with accurate FTICR is shown be capable characterizing mixtures significantly more than 105 components accuracies < 1 ppm, sufficient broad identification AMTs. Attractions approach include capability automated confidence identification, unbiased proteome coverage, exploiting stable-isotope labeling methods realize precision relative abundance measurements, potential study mammalian proteomes when additional sample fractionation. demonstrated selected examples Saccharomyces cerevisiae, Deinococcus radiodurans, mouse melanoma cells.