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Isoform-Specific Quantitative Proteomics Applying N-terminal Enrichment and informatics deconvolution


EMSL Project ID
29412

Abstract

For higher eukaryotic organisms, a single gene can produce multiple protein products (or isoforms) by various mechanisms, such as alternative splicing and post-translational proteolytic processing. Quantitative proteomics have been widely applied for studying protein abundance changes based on peptide-level measurements using a "bottom-up" strategy; however, the general challenge of this "bottom-up" approach is that it is unable to resolve protein isoforms. The quantitative measurement of specific protein products or isoforms that participate in a functional network may be essential for deciphering cell signaling and could represent the next era of proteomics.

This project is directed towards developing a protein N-termini enrichment strategy for quantitative analysis of isoform-specific protein abundance changes; in particular, proteolytic isoforms. The N-termini subproteome and global proteome will be quantitatively analyzed with the isoform-specific quantitative information eventually revealed by informatics deconvolution. Initially, we will focus on the development and optimization of an efficient sample preparation method to specifically enrich the N-terminal peptides. Once the enrichment approach is optimized, it will be coupled to quantitative strategies and demonstrated in different applications.

Project Details

Start Date
2008-04-01
End Date
2009-04-05
Status
Closed

Team

Principal Investigator

Weijun Qian
Institution
Pacific Northwest National Laboratory

Team Members

Brianne Petritis
Institution
Pacific Northwest National Laboratory