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Genomes-to-Life Protein Complex Isolation Capability


EMSL Project ID
6504a

Abstract

We have established a pilot proteomics facility. The primary aim of this facility is to develop new approaches for rapid, robust isolation of protein complexes as well as develop methods to analyze protein complexes, including a determination of their component stoichiometry, modifications, and relative levels. An important part of this research program is the integration of computational tools that will combine experimental information with proteomics data. Two primary approaches are being taken to isolate protein complexes: 1) expression of epitope-tagged proteins in target cells to allow direct isolation, 2) generation of affinity reagents using yeast surface display technology to allow isolation of native complexes. To date, we have cloned and expressed over 20 proteins from Shewanella oneidensis to use in our initial screenings and have built an appropriate bioinformatics infrastructure to track sample handling and manage the large amounts of data generated by this project. We are also exploring high-throughput, automated approaches to sample preparation, cross-linking and mass spectrometry analysis.

Project Details

Start Date
2007-01-17
End Date
2009-03-15
Status
Closed

Team

Principal Investigator

Brian Hooker
Institution
Pacific Northwest National Laboratory

Team Members

Chiann-tso Lin
Institution
Pacific Northwest National Laboratory

H Wiley
Institution
Environmental Molecular Sciences Laboratory