Discrimination between stable and dynamic components of protein complexes by means of quantitative proteomics.
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.
Kito
Keiji
K
Department of Computational Biology, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Japan.
Kawaguchi
Noriko
N
Okada
Satoshi
S
Ito
Takashi
T
D017489:CDC28 Protein Kinase, S cerevisiae / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid; D016213:Cyclins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D015852:Eukaryotic Initiation Factor-2 / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D020717:Eukaryotic Initiation Factor-2B / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D007553:Isotope Labeling; D013058:Mass Spectrometry; D008954:Models, Biological; D011485:Protein Binding; D021122:Protein Subunits / Q000378:metabolism; D040901:Proteomics; D015203:Reproducibility of Results; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
D016213:Cyclins; D015852:Eukaryotic Initiation Factor-2; D020717:Eukaryotic Initiation Factor-2B; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D017489:CDC28 Protein Kinase, S cerevisiae