Oligomerization-function relationship of EGFR on living cells detected by the coiled-coil labeling and FRET microscopy

Biochimica Et Biophysica Acta
Hirotaka YamashitaKatsumi Matsuzaki

Abstract

The epidermal growth factor receptor (EGFR) is a well-studied receptor tyrosine kinase and an important anticancer therapeutic target. The activity of EGFR autophosphorylation and transphosphorylation, which induces several cell signaling pathways, has been suggested to be related to its oligomeric state. However, the oligomeric states of EGFRs induced by EGF binding and the receptor-ligand stoichiometry required for its activation are still controversial. In the present study, we performed Förster resonance energy transfer (FRET) measurements by combining the coiled-coil tag-probe labeling method and spectral imaging to quantitatively analyze EGFR oligomerization on living CHO-K1 cell membranes at physiological expression levels. In the absence of its ligands, EGFRs mainly existed as monomers with a small fraction of predimers (~10%), whereas ~70% of the EGFRs formed dimers after being stimulated with the ligand EGF. Ligand-induced dimerization was not significantly affected by the perturbation of membrane components (cholesterol or monosialoganglioside GM3). We also investigated both dose and time dependences of EGF-dependent EGFR dimerization and autophosphorylation. The formation of dimers occurred within 20s of the ligand ...Continue Reading

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Citations

Mar 11, 2016·Molecular BioSystems·Jonathan LotzeAnnette G Beck-Sickinger
Aug 10, 2018·Chemistry : a European Journal·W Mathis Rink, Franziska Thomas
Nov 26, 2015·The Journal of Biological Chemistry·Mi-Jung KwonEok-Soo Oh
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May 3, 2021·Biochimica Et Biophysica Acta. Biomembranes·Mikito NakanoMichio Murata

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