The Dynamics and Localization of the Epidermal Growth Factor Receptor (EGFR) on Live Cell Plasma Membrane Studied by ITIR-FCS

摘要: The Epidermal Growth Factor Receptor (EGFR), a prototypical receptor tyrosine kinase, plays an important role in cellular processes such as proliferation, migration and apoptosis. EGFR is known to be strongly regulated by its localization the state of activation on plasma membrane. However, spatial distribution membrane has not been resolved detail.Here, we use Imaging Total Internal Reflection Fluorescence Correlation Spectroscopy (ITIR-FCS) investigate behavior live cells before after stimulation. ITIR-FCS allows monitoring dynamics cell membranes combination with FCS diffusion laws provides information nanoscopic organization.Our results indicate that partly trapped cholesterol sensitive domains also couples cytoskeleton. Stimulation EGF activates dose dependent manner. At low-dose stimulation (10 ng/ml), confinement remain unchanged internalization independent recycles back high-dose treatment (100 endocytosis cholesterol-depletion impairs internalization. In addition, concomitant leads stronger receptor, dose, clusters accumulate Furthermore, actin cytoskeleton promotes ligand Disruption inhibits induces clustering This study demonstrates capabilities follow events cells. It only but pathway dependent, thus providing new insights into organization