Spatial and Temporal Proteomics
Pacific Northwest National Laboratory's (PNNL's) Spatial and Temporal Proteomics research team is working to understand how self-organizing signaling networks integrate competing environmental stimuli to promote a range of different cellular responses that underlie adaptive and pathological behavior. Currently, it is clear that adaptive cellular responses to environmental signals frequently act through the modulation of specific protein complexes at the cell surface, which modulate protein trafficking and the protein composition of the plasma membrane and other organelles. The molecular triggers associated with cellular reprogramming involve both post-translational modifications, such as phosphorylation, and changes in protein abundance.
Our hypothesis is that the identification of site-specific, post-translational modifications and associated changes in the composition and protein-protein associations within individual organelles will reveal both the central nodes of signal transduction cascades and their cellular integration. However, current methodologies to isolate cellular organelles and to identify their protein composition and associated post-translational modifications are inadequate to permit an unbiased assessment of signaling mechanisms. Therefore, we are building upon the technological expertise in proteomics already existing at PNNL by developing affinity methods for the simultaneous isolation of multiple organelles. We are also developing complementary separation and mass spectrometric methods that will permit identification of the protein compositional changes and post-translational modifications within individual organelles.
Three of these projects focus on protein imaging and affinity methods for organelle isolation.
- High-Throughput Screening of Protein Localization
- High-Throughput Isolation of Organelles
- Analysis of Protein Function in Living Cells
Four projects are focused on mass spectrometric protein identification.
- Quantitative Characterization of Post-Translational Protein Modifications Using Mass Spectrometry
- Fast, Two-Dimensional, Gas-Phase Separations for Ultrahigh-Throughput Global Analyses
- Proteomics of Membrane Proteins: Relating Calcium Signaling and Oxidative Stress
- Functional Genomic Analysis of the Regulation of Bone Cells by a Bioactive Lipid