Visualization of EGFR and Grb2 Interaction Using Bimolecular Fluorescence Complementation

摘要

The dimerization of epidermal growth factor receptor (EGFR) and its endocytic transport are important in regulating signal transduction. In the present study, we applied the strategy of Bimolecular Fluorescence Complementation (BiFC), EGFR homodimer and hetrodimer of EGFR and its partner Grb2 in cells were visualized. This strategy takes advantage of the ability of two nonfluorescent fragments of Venus fluorescent protein to form a fluorescent signal when fused to the amino termini of EGFR and Grb2. Since EGFR is a large protein contains more than 1800 amido acids, proper fold of the fusion protein is essential for the BiFC assay of EGFR and its partners. Our results indicate that BiFC is a suitable application for research of EGFR interaction with other proteins.