Luciferase fragment complementation imaging of conformational changes in the epidermal growth factor receptor.

The Journal of Biological Chemistry
Katherine S YangLinda J Pike

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.

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Citations

Dec 23, 2011·Proceedings of the National Academy of Sciences of the United States of America·Jennifer L Macdonald-ObermannLinda J Pike
Jul 24, 2012·The Journal of Biological Chemistry·Yu LiLinda J Pike
Jul 9, 2014·Analytical and Bioanalytical Chemistry·Taha AzadSaman Hosseinkhani
May 12, 2010·Chemical Biology & Drug Design·Robert Y C YangGarland R Marshall
Aug 5, 2015·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Ichiro N Maruyama
Jun 23, 2011·Analytical Biochemistry·Amjad P KhanAlnawaz Rehemtulla
Jan 27, 2015·Annual Review of Biochemistry·Erika KovacsJohn Kuriyan
Sep 5, 2014·Molecular Endocrinology·Ying LiuStuart J Frank
Sep 10, 2013·The Journal of Biological Chemistry·Jennifer L Macdonald-ObermannLinda J Pike
Nov 20, 2019·European Biophysics Journal : EBJ·Noga Kozer, Andrew H A Clayton
Jan 17, 2015·Oncoscience·Madryn C Lake, Eric O Aboagye
Nov 16, 2012·Scientific Reports·Lijuan He, Kalina Hristova

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