title: High-throughput comparative proteome analysis using a quantitative cysteinyl-peptide enrichment technology.
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.
authors.affiliation: Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, P.O. Box 999, Richland, Washington 99352, USA.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.forename: Wei-Jun
authors.initials: WJ
authors.lastname: Qian
authors.forename: Eric F
authors.initials: EF
authors.lastname: Strittmatter
authors.forename: David G
authors.initials: DG
authors.lastname: Camp
authors.forename: Gordon A
authors.initials: GA
authors.lastname: Anderson
authors.forename: Brian D
authors.initials: BD
authors.lastname: Thrall
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
mesh_terms: D002478:Cells, Cultured; D003545:Cysteine / Q000737:chemistry*; D006801:Humans; D010455:Peptides / Q000032:analysis* / Q000737:chemistry*; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*
chemical_list: D010455:Peptides; D020543:Proteome; D003545:Cysteine