Characterization of the EGFR interactome reveals associated protein complex networks and intracellular receptor dynamics

Abstract

Growth factor receptor mediated signaling is meanwhile recognized as a complex signaling network, which is initiated by recruiting specific patterns of adaptor proteins to the intracellular domain of epidermal growth factor receptor (EGFR). Approaches to globally identify EGFR-binding proteins are required to elucidate this network. We affinity-purified EGFR with its interacting proteins by coprecipitation from lysates of A431 cells. A total of 183 proteins were repeatedly detected in high-resolution MS measurements. For 15 of these, direct interactions with EGFR were listed in the iRefIndex interaction database, including Grb2, shc-1, SOS1 and 2, STAT 1 and 3, AP2, UBS3B, and ERRFI. The newly developed Cytoscape plugin ModuleGraph allowed retrieving and visualizing 93 well-described protein complexes that contained at least one of the proteins found to interact with EGFR in our experiments. Abundances of 14 proteins were modulated more than twofold upon EGFR activation whereof clathrin-associated adaptor complex AP-2 showed 4.6-fold enrichment. These proteins were further annotated with different cellular compartments. Finally, interactions of AP-2 proteins and the newly discovered interaction of CIP2A could be verified. In conclusion, a powerful technique is presented that allowed identification and quantitative assessment of the EGFR interactome to provide further insight into EGFR signaling.

A Kinase-Independent Role for EGF Receptor in Autophagy Initiation

“…Recently, exocyst complexes have been implicated in starvation induced autophagy (Bodemann et al, 2011). A proteomic screen demonstrated that exocyst subunits interact with the EGFR complex (Foerster et al, 2013); we therefore explored if LAPTM4B and inactive EGFR interact with exocyst complex to modulate autophagy. As shown in Figures 5A and 5B, LAPTM4B and EGFR interact with multiple subunits of the exocyst complex.…”

IQGAPs choreograph cellular signaling from the membrane to the nucleus

“…Signal transduction from the receptor at the plasma membrane to downstream MAPK components requires coordination among signaling proteins. IQGAP1 associates with several MAPK proteins, including EGFR [42, 43], K-Ras [44], Raf [45, 46], MEK [14] and ERK [13, 14] (Figure 1Ai) (Table 1), providing a scaffold which promotes EGF-induced MAPK signaling [13, 45]. IQGAP3, which interacts with H-Ras [47] and ERK1 [48], also augments EGF-stimulated MAPK signaling.…”

Cohort profile: Greifswald approach to individualized medicine (GANI_MED)

“…The interactome of the epidermal growth factor receptor (EGFR) was identified and quantified. The newly developed Cytoscape plugin ModuleGraph facilitated the extension and functional investigation of this network [74]. A short 5-item questionnaire for routine clinical assessment of childhood abuse and neglect has been developed and validated [3,75].…”