Quantitative Proteome Analysis of Breast Cancer Cell Lines using 18O-Labeling and an Accurate Mass and Time Tag Strategy
Proteome comparison of cell lines derived from breast cancer and normal breast epithelium provide opportunities to identify differentially expressed proteins and pathways associated with specific phenotypes. We employed trypsin-catalyzed 16O/18O peptide labeling, FTI-CR mass spectrometry, and the accurate mass and time (AMT) tag strategy to calculate compare the relative protein abundances of hundreds of proteins simultaneously in non-cancer and cancer cell lines derived from breast tissue. A reference panel of cell lines was created to facilitate comparisons of relative protein abundance amongst multiple cell lines and across multiple experiments. A peptide database generated from multidimensional LC separations and MS/MS analysis was used to facilitate subsequent AMT tag-based peptide identifications. This peptide database represented a total of 2,299 proteins, including 514 that were quantified using the AMT tag and 16O/18O strategies. Eighty-six proteins showed at least a 3-fold protein abundance change between cancer and non-cancer cell lines. A comparison of protein expression profiles with previously published gene expression data revealed that 21 of these proteins also had >3-fold differences between the non-cancer and cancer cell lines at the transcriptional level. Clustering of protein abundance ratios revealed that several groups of proteins were differentially expressed between the cancer cell lines
- Research Organization:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 885447
- Report Number(s):
- PNNL-SA-46350; 2625; 400412000
- Journal Information:
- Proteomics, 6(9):2903-2915, Journal Name: Proteomics, 6(9):2903-2915 Journal Issue: 9 Vol. 6
- Country of Publication:
- United States
- Language:
- English
Similar Records
Proteomic Analyses using an Accurate Mass and Time Tag Strategy
The Use of Accurate Mass Tags for High-Throughput Microbial Proteomics