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Luciferase Fragment Complementation Imaging of Conformational Changes in the Epidermal Growth Factor Receptor
- Source :
- Journal of Biological Chemistry. 284:7474-7482
- Publication Year :
- 2009
- Publisher :
- Elsevier BV, 2009.
-
Abstract
- 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.
- Subjects :
- Recombinant Fusion Proteins
CHO Cells
Biology
Biochemistry
Tropomyosin receptor kinase C
Mice
Cricetulus
Growth factor receptor
Epidermal growth factor
Cricetinae
Enzyme-linked receptor
Animals
Luciferase
Epidermal growth factor receptor
Luciferases
Molecular Biology
Insulin-like growth factor 1 receptor
Mitogen-Activated Protein Kinase Kinases
Epidermal Growth Factor
Mechanisms of Signal Transduction
Cell Biology
Protein Structure, Tertiary
Cell biology
ErbB Receptors
biology.protein
Dimerization
Tyrosine kinase
hormones, hormone substitutes, and hormone antagonists
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 284
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....b28253c9c7553a2a87799de34e809fea
- Full Text :
- https://doi.org/10.1074/jbc.m808041200