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A New Approach for the Comparative Analysis of Multiprotein Complexes Based on 15N Metabolic Labeling and Quantitative Mass Spectrometry
Published on: March 13, 2014
Discrimination between stable and dynamic components of protein complexes by means of quantitative proteomics
Keiji Kito1, Noriko Kawaguchi, Satoshi Okada
1Department of Computational Biology, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Japan.
Abstract:
To discriminate between stable and dynamic protein-protein interactions, we propose a strategy in which cells with and without tagged bait are differentially labeled with stable isotope and combined prior to complex purification. Mass-spectrometric analysis of the purified complexes identifies stable and dynamic components as those derived exclusively from the tagged cells and those from both cells, respectively. We successfully applied this strategy to analyze two yeast protein complexes, eIF2B-eIF2 and cyclin-Cdc28.
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