Proteomic analysis of scaffold proteins in the ERK cascade.
ERK cascade scaffolds serve as docking platforms to coordinate the assembly of multiprotein complexes that contribute to the spatial and temporal control of ERK signaling. Given that protein-protein interactions are essential for scaffold function, determining the full repertoire of scaffold binding partners will likely provide new insight into the regulation and activities of the ERK cascade scaffolds. In this chapter, we describe methods to identify scaffold interacting proteins using a proteomics approach. This protocol is based on the affinity purification of scaffold complexes from tissue culture cells and utilizes mass spectrometry to identify the protein constituents of the complex.
McKay
Melissa M
MM
Laboratory of Cell and Developmental Signaling, Center for Cancer Research, National Cancer Institute-Frederick, Frederick, MD, USA.
Morrison
Deborah K
DK
D048049:Extracellular Signal-Regulated MAP Kinases / Q000302:isolation & purification / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D020935:MAP Kinase Signaling System; D013058:Mass Spectrometry; D040901:Proteomics / Q000379:methods*; D012117:Resins, Synthetic
D012117:Resins, Synthetic; D048049:Extracellular Signal-Regulated MAP Kinases
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