期刊
PROTEOMICS CLINICAL APPLICATIONS
卷 5, 期 7-8, 页码 383-396出版社
WILEY-BLACKWELL
DOI: 10.1002/prca.201000115
关键词
Cancer biology; LC-MRM; Pathways; Quantification; Signaling
资金
- US Army Medical Research and Materiel Command [DAMD17-02-2-0051]
- National Functional Genomics Center
- National Cancer Institute [P30-CA076292, R21CA141285-01, RO1CA077859]
- Moffitt Foundation
- Florida Department of Health [06BS-02-9614]
- University of Florida-Moffitt Collaborative Partnership
- Department of Defense
- Melanoma Research Foundation
- Bankhead-Coley Research Program of the State of Florida [09BN-14]
- American Cancer Society [93-032-13]
- NIH/National Cancer Institute [U54 CA143970-01]
- [W81XWH-08-2-0101]
Purpose: The Quantitative Assay Database (QuAD), http://proteome.moffitt.org/QUAD/, facilitates widespread implementation of quantitative mass spectrometry in cancer biology and clinical research through sharing of methods and reagents for monitoring protein expression and modification. Experimental design: Liquid chromatography coupled to multiple reaction monitoring (LC-MRM) mass spectrometry assays are developed using SDS-PAGE fractionated lysates from cancer cell lines. Pathway maps created using GeneGO Metacore provide the biological relationships between proteins and illustrate concepts for multiplexed analysis; each protein can be selected to examine assay development at the protein and peptide levels. Results: The coupling of SDS-PAGE and multiple reaction monitoring mass spectrometry screening has been used to detect 876 peptides from 218 cancer-related proteins in model systems including colon, lung, melanoma, leukemias, and myeloma, which has led to the development of 95 quantitative assays including stable-isotope-labeled peptide standards. Methods are published online and peptide standards are made available to the research community. Protein expression measurements for heat shock proteins, including a comparison with ELISA and monitoring response to the HSP90 inhibitor, 17-(dimethyl-aminoethylamino)-17-demethoxygeldanamycin (17-DMAG), are used to illustrate the components of the QuAD and its potential utility. Conclusions and Clinical Relevance: This resource enables quantitative assessment of protein components of signaling pathways and biological processes and holds promise for systematic investigation of treatment responses in cancer.
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