High-throughput comparative proteome analysis using a quantitative cysteinyl-peptide enrichment technology.
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.
Liu
Tao
T
Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, P.O. Box 999, Richland, Washington 99352, USA.
Qian
Wei-Jun
WJ
Strittmatter
Eric F
EF
Camp
David G
DG
Anderson
Gordon A
GA
Thrall
Brian D
BD
Smith
Richard D
RD
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*
D010455:Peptides; D020543:Proteome; D003545:Cysteine