High-Throughput Comparative Proteome Analysis Using a Quantitative Cysteinyl-peptide Enrichment Technology
- 5 August 2004
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 76 (18) , 5345-5353
- https://doi.org/10.1021/ac049485q
Abstract
A new quantitative cysteinyl-peptide enrichment technology (QCET) was developed to achieve higher efficiency, greater dynamic range, and higher throughput in quantitative proteomics that use stable-isotope labeling techniques combined with high-resolution liquid chromatography (LC)-mass spectrometry (MS). This approach involves (18)O labeling of tryptic peptides, high-efficiency enrichment of cysteine-containing peptides, and confident protein identification and quantification using the accurate mass and time tag strategy. Proteome profiling of naïve and in vitro-differentiated human mammary epithelial cells using QCET resulted in the identification and quantification of 603 proteins in a single LC-Fourier transform ion cyclotron resonance MS analysis. Advantages of this technology include the following: (1) a simple, highly efficient method for enriching cysteinyl-peptides; (2) a high-throughput strategy suitable for extensive proteome analysis; and (3) improved labeling efficiency for better quantitative measurements. This technology enhances both the functional analysis of biological systems and the detection of potential clinical biomarkers.Keywords
This publication has 13 references indexed in Scilit:
- An automated high performance capillary liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometer for high-throughput proteomicsJournal of the American Society for Mass Spectrometry, 2004
- Improved β-Elimination-Based Affinity Purification Strategy for Enrichment of PhosphopeptidesAnalytical Chemistry, 2003
- Proteome analyses using accurate mass and elution time peptide tags with capillary LC time-of-flight mass spectrometryJournal of the American Society for Mass Spectrometry, 2003
- Trypsin catalyzed 16O-to-18O exchange for comparative proteomics: Tandem mass spectrometry comparison using MALDI-TOF, ESI-QTOF, and ESI-ion trap mass spectrometersJournal of the American Society for Mass Spectrometry, 2003
- Mass Spectrometric Analysis of Protein Mixtures at Low Levels Using Cleavable 13C-Isotope-coded Affinity Tag and Multidimensional ChromatographyMolecular & Cellular Proteomics, 2003
- Use of Artificial Neural Networks for the Accurate Prediction of Peptide Liquid Chromatography Elution Times in Proteome AnalysesAnalytical Chemistry, 2003
- Global analysis of the Deinococcus radiodurans proteome by using accurate mass tagsProceedings of the National Academy of Sciences, 2002
- Analysis of Quantitative Proteomic Data Generated via Multidimensional Protein Identification TechnologyAnalytical Chemistry, 2002
- Minimizing Resolution of Isotopically Coded Peptides in Comparative ProteomicsJournal of Proteome Research, 2002
- Proteome analysis using selective incorporation of isotopically labeled amino acidsJournal of the American Society for Mass Spectrometry, 2000