We propose to develop an approach to allow rapid identification of proteins in complex mixtures suitable for protein expression mapping in total cell lysates. A robust approach to identify proteins as mixtures will advance cancer research in several areas. First, the approach has the potential to significantly improve the sensitivity of analysis of proteins. Benefits to cancer research will derive from the ability to identify proteins using smaller numbers of cells leading to direct examination of tumor biopsy samples. Rigorous molecular analysis at the protein level, encompassing separation, visualization, and identification, is currently difficult for small quantities of protein. The methodology proposed is general and therefore can be applied to the monitoring of gene produce expression, analysis and detection of cellular localization and post translational modification of proteins, and monitoring of major signal transduction networks involved in cancer. Second, the approach has great potential for automation allowing the technique to be transferred to laboratories involved in cancer research eliminating the need to collaborate with experts. Last, by developing the ability to quantitate and perform subtractive analyses changes in protein expression related to neoplastic transformation could be observed.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants Phase II (R33)
Project #
5R33CA081665-03
Application #
6174107
Study Section
Special Emphasis Panel (ZCA1-SRRB-C (J2))
Program Officer
Gallahan, Daniel L
Project Start
1999-04-01
Project End
2002-03-31
Budget Start
2000-04-13
Budget End
2001-03-31
Support Year
3
Fiscal Year
2000
Total Cost
$281,981
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Pebernard, Stephanie; McDonald, W Hayes; Pavlova, Yelena et al. (2004) Nse1, Nse2, and a novel subunit of the Smc5-Smc6 complex, Nse3, play a crucial role in meiosis. Mol Biol Cell 15:4866-76
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Tabb, David L; Smith, Lori L; Breci, Linda A et al. (2003) Statistical characterization of ion trap tandem mass spectra from doubly charged tryptic peptides. Anal Chem 75:1155-63
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Wu, Christine C; MacCoss, Michael J; Howell, Kathryn E et al. (2003) A method for the comprehensive proteomic analysis of membrane proteins. Nat Biotechnol 21:532-8
McDonald, W Hayes; Yates 3rd, John R (2003) Shotgun proteomics: integrating technologies to answer biological questions. Curr Opin Mol Ther 5:302-9

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