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Protein Complex Identification using novel affinity resins


EMSL Project ID
12091a

Abstract

We are validating the use of a FlAsH resin with a dithiol releasing agent for high-throughput protein complex identification We are going to do endogenous pull-downs of protein complexes from S. oneidensis and validations in E. coli and would like to use LCMS-MS/SEQUEST to identify the proteins in our complex mixtures. We will be submitting trypsinized peptides from protein mixtures, in which 2-20 proteins should be highly enriched. We expect that in terms of mass spectrometry these samples will be well-suited to the classic Shewanella federation run times and approaches. In total, we estimate between 60 and 150 samples in this fiscal year.

Project Details

Start Date
2006-11-07
End Date
2009-11-08
Status
Closed

Team

Principal Investigator

M. Mayer-Cumblidge
Institution
Pacific Northwest National Laboratory

Team Members

Ping Yan
Institution
Pacific Northwest National Laboratory

Liang Shi
Institution
Pacific Northwest National Laboratory

Thomas Squier
Institution
Western University of Health Sciences

Related Publications

Yan P, T Wang, GJ Newton, TV Knyushko, Y Xiong, DJ Bigelow, TC Squier, and MU Mayer. 2009. "A Targeted Releasable Affinity Probe (TRAP) for In Vivo Photo-Crosslinking." Chembiochem 10(9):1507-1518. doi:10.1002/cbic.200900029