Crystal structures of the epidermal growth factor (EGF) receptor suggest that its activation is associated with extensive conformational changes in both the extracellular and intracellular domains. However, evidence of these structural dynamics in intact cells has been lacking. Here we use luciferase complementation imaging to follow EGF-induced conformational changes in its receptor in real time in live cells. When the luciferase fragments are fused to the C terminus of an EGF receptor lacking the cytoplasmic domain, EGF stimulates a rapid increase in luciferase activity, consistent with ligand-induced receptor dimerization. However, when the luciferase fragments are fused to the C terminus of the full-length receptor, EGF induces a rapid but transient decrease in luciferase activity. The decrease requires tyrosine kinase activity, whereas the subsequent recovery requires MAP kinase activity. Our data demonstrate the utility of the luciferase system for in vivo imaging changes in EGF receptor dimerization and conformation. They also identify two sequential ligand-induced conformational changes in the EGF receptor.
Epidermal growth factor (EGF) receptor T669 peptide kinase from 3T3-L1 cells is an EGF-stimulated "MAP" kinase
Epidermal growth factor induces rapid, reversible aggregation of the purified epidermal growth factor receptor
A mutant epidermal growth factor receptor with defective protein tyrosine kinase is unable to stimulate proto-oncogene expression and DNA synthesis.
Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells
Unliganded epidermal growth factor receptor dimerization induced by direct interaction of quinazolines with the ATP binding site.
Preformed oligomeric epidermal growth factor receptors undergo an ectodomain structure change during signaling
Ligand-independent dimer formation of epidermal growth factor receptor (EGFR) is a step separable from ligand-induced EGFR signaling
Structure of the epidermal growth factor receptor kinase domain alone and in complex with a 4-anilinoquinazoline inhibitor.
Crystal structure of a truncated epidermal growth factor receptor extracellular domain bound to transforming growth factor alpha
Crystal structure of the complex of human epidermal growth factor and receptor extracellular domains
Kinetics of regulated protein-protein interactions revealed with firefly luciferase complementation imaging in cells and living animals
A unique structure for epidermal growth factor receptor bound to GW572016 (Lapatinib): relationships among protein conformation, inhibitor off-rate, and receptor activity in tumor cells
Ligand-induced dimer-tetramer transition during the activation of the cell surface epidermal growth factor receptor-A multidimensional microscopy analysis
Conformational changes accompany phosphorylation of the epidermal growth factor receptor C-terminal domain
The membrane proximal disulfides of the EGF receptor extracellular domain are required for high affinity binding and signal transduction but do not play a role in the localization of the receptor to lipid rafts
The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor AG1478 increases the formation of inactive untethered EGFR dimers. Implications for combination therapy with monoclonal antibody 806
A system for quantifying dynamic protein interactions defines a role for Herceptin in modulating ErbB2 interactions
Investigation of the dimerization of proteins from the epidermal growth factor receptor family by single wavelength fluorescence cross-correlation spectroscopy
Oligomerization of the EGF receptor investigated by live cell fluorescence intensity distribution analysis
Noninvasive imaging and quantification of epidermal growth factor receptor kinase activation in vivo.
ERK-dependent threonine phosphorylation of EGF receptor modulates receptor downregulation and signaling.
Mechanics of EGF receptor/ErbB2 kinase activation revealed by luciferase fragment complementation imaging
The intracellular juxtamembrane domain of the epidermal growth factor (EGF) receptor is responsible for the allosteric regulation of EGF binding.
Asp-960/Glu-961 controls the movement of the C-terminal tail of the epidermal growth factor receptor to regulate asymmetric dimer formation.
Development of a novel molecular sensor for imaging estrogen receptor-coactivator protein-protein interactions.
A versatile platform to analyze low-affinity and transient protein-protein interactions in living cells in real time
Identification of function-regulating antibodies targeting the receptor protein tyrosine phosphatase sigma ectodomain
Epidermal growth factor receptors containing a single tyrosine in their C-terminal tail bind different effector molecules and are signaling-competent
Sequence-specific 5mC detection in live cells based on the TALE-split luciferase complementation system
Dynamic analysis of the epidermal growth factor (EGF) receptor-ErbB2-ErbB3 protein network by luciferase fragment complementation imaging.
Different epidermal growth factor (EGF) receptor ligands show distinct kinetics and biased or partial agonism for homodimer and heterodimer formation.
Different Epidermal Growth Factor Receptor (EGFR) Agonists Produce Unique Signatures for the Recruitment of Downstream Signaling Proteins.
The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.