European Biophysics Journal

, Volume 37, Issue 4, pp 469–481 | Cite as

Studies of distribution, location and dynamic properties of EGFR on the cell surface measured by image correlation spectroscopy

Original Paper

Abstract

In this work, we have studied the distribution and dynamic properties of Epidermal Growth Factor (EGF) receptors in the plasma membrane of fixed and live cells as well as the extent of co-localization of this transmembrane protein with proteins specific for three-membrane microdomains: membrane rafts, caveolae and clathrin-coated pits. This was achieved using a family of image-processing tools called image correlation spectroscopy (ICS), image cross-correlation spectroscopy (ICCS) and dynamic image correlation spectroscopy (DICS). Our results indicate that EGFR is diffusely distributed on the cell surface at 37°C and aggregates as the temperature is lowered to 4°C. This aggregation takes place within 15 min and is reversible. Changes in temperature also affect the diffusion of EGFR by two orders of magnitude. The dynamic properties of EGFR are similar to the dynamic properties of a GPI-anchored protein known to be present in membrane rafts, which motivated us to explore the extent of co-localization of EGFR with this membrane raft protein using ICCS. Our results indicate that more than half of the EGFR population is present in membrane rafts and smaller percentages are present in caveolae and clathrin-coated pits.

Keywords

EGFR Diffusion Plasma membrane microdomains Membrane rafts Temperature dependence Image correlation spectroscopy 

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Copyright information

© EBSA 2007

Authors and Affiliations

  • Eleonora Keating
    • 1
  • Anja Nohe
    • 2
  • Nils O. Petersen
    • 1
    • 3
  1. 1.Department of ChemistryThe University of Western OntarioLondonCanada
  2. 2.Department of Chemical Engineering and Institute for Molecular BiophysicsUniversity of MaineOronoUSA
  3. 3.National Institute for NanotechnologyNational Research Council CanadaEdmontonCanada

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