Nanoscale imaging of epidermal growth factor receptor clustering: effects of inhibitors.
The development of some solid tumors is associated with overexpression of the epidermal growth factor receptor (EGFR) and often correlates with poor prognosis. Near field scanning optical microscopy, a technique with subdiffraction-limited optical resolution, was used to examine the influence of two inhibitors (the chimeric 225 antibody and tyrosine phosphorylation inhibitor AG1478) on the nanoscale clustering of EGFR in HeLa cells. The EGFR is organized in small clusters, average diameter of 150 nm, on the plasma membrane for both control and EGF-treated cells. The numbers of receptors in individual clusters vary from as few as one or two proteins to greater than 100. Both inhibitors yield an increased cluster density and an increase in the fraction of clusters with smaller diameters and fewer receptors. Exposure to AG1478 also decreases the fraction of EGFR that colocalizes with both rafts and caveolae. EGF stimulation results in a significant loss of the full-length EGFR from the plasma membrane with the concomitant appearance of low molecular mass proteolytic products. By contrast, AG1478 reduces the level of EGFR degradation. Changes in receptor clustering provide one mechanism for regulating EGFR signaling and are relevant to the design of strategies for therapeutic interventions based on modulating EGFR signaling.
Abulrob
Abedelnasser
A
Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario K1A 0R5, Canada. Abedelnasser.Abulrob@nrc.gc.ca
Lu
Zhengfang
Z
Baumann
Ewa
E
Vobornik
Dusan
D
Taylor
Rod
R
Stanimirovic
Danica
D
Johnston
Linda J
LJ
D002462:Cell Membrane / Q000378:metabolism; D019281:Dimerization; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism; D008853:Microscopy / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D036103:Nanotechnology / Q000379:methods; D011379:Prognosis; D017434:Protein Structure, Tertiary; D011799:Quinazolines; D020032:Tyrphostins / Q000494:pharmacology
D004791:Enzyme Inhibitors; D011799:Quinazolines; D020032:Tyrphostins; C101044:RTKI cpd; D066246:ErbB Receptors