An integrated workflow for identification and quantitation of intact phosphoproteins
EMSL Project ID
34504
Abstract
Multisite protein modifications involving phosphorylation alone or in combination with other post-translational modifications (PTMs) play an essential role in regulating protein activity. Hence, knowing the overall modification signature of proteins is essential to gain a more complete understanding of the complex signaling mechanisms that regulate responses to environmental stimuli and that impact human health. Given the limitations of conventional (bottom up) proteomics, there has been increasing interest in top down proteome characterization strategies, where individual proteins are selected for analysis by tandem MS, without the need for prior chemical or enzymatic proteolysis. When applied to complex mixtures of proteins, these strategies presently suffer from limited sensitivity and throughput, are best suited for small proteins, and cannot accurately detect phosphorylation, a major regulatory PTM. The main goals of this proposal are to develop a technology platform to quantitatively map multiple PTMs (with emphasis on phosphorylation) occurring within protein complexes with high sensitivity and throughput and test the hypothesis that this system can be used to identify protein signatures associated with specific biological responses.
Project Details
Start Date
2009-04-28
End Date
2012-04-29
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Wu S, N Tolic, Z Tian, EW Robinson, and L Pasa-Tolic. 2011. "An integrated top-down and bottom-up strategy for characterization protein isoforms and modifications." Chapter 18 in Bioinformatics for Comparative Proteomics: Methods in Molecular Biology, vol. 694, ed. CH Wu and C Chen, pp. 291-304. Springer, New York, NY.