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Protein Complex Identification using novel affinity resins


EMSL Project ID
12091

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 would like to use LCQ/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 100 and 400 samples in this fiscal year.

Project Details

Project type
Exploratory Research
Start Date
2005-02-24
End Date
2007-02-27
Status
Closed

Team

Principal Investigator

M. Mayer-Cumblidge
Institution
Pacific Northwest National Laboratory

Team Members

Liang Shi
Institution
Pacific Northwest National Laboratory

Thomas Squier
Institution
Western University of Health Sciences

Related Publications

B Chen, TC Squier, MUljana Mayer, and Cao H. 2006. "CrAsH: A Biarsenical Multi-use Affinity Probe with low non-specific fluorescence." Chemical Communications(24):2601-2603.
Verma S, Xiong Y, Mayer MU, Squier TC (2007) Remodeling of the Bacterial RNA Polymerase Supramolecular Complex in Response to Environmental Conditions. Biochemistry 46: 3023-3035
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