Resolving the EGF-EGFR interaction characteristics through a multiple-temperature, multiple-inhibitor, real-time interaction analysis approach

  • Authors:
    • Hanna Björkelund
    • Lars Gedda
    • Magnus Malmqvist
    • Karl Andersson
  • View Affiliations

  • Published online on: October 30, 2012     https://doi.org/10.3892/mco.2012.37
  • Pages: 343-352
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Abstract

Overexpression and aberrant activity of the epidermal growth factor (EGF) have been observed in various cancer types, rendering it an important target in oncology research. The interaction between EGF and its receptor (EGFR), as well as subsequent internalization, is complex and may be affected by various factors including tyrosine kinase inhibitors (TKIs). By combining real‑time binding curves produced in LigandTracer® with internalization assays conducted at different temperatures and with different TKIs, the processes of ligand binding, internalization and excretion was visualized. SKOV3 cells had a slower excretion rate compared to A431 and U343 cells, and the tested TKIs (gefitinib, lapatinib, AG1478 and erlotinib) reduced the degree of internalization. The kinetic analysis of the binding curves further demonstrated TKI‑dependent balances of EGFR monomer and dimer populations, where lapatinib promoted the monomeric form, while the other TKIs induced dimers. The dimer levels were found to be associated with the apparent affinity of the EGF‑EGFR interaction, with EGF binding stronger to EGFR dimers compared to monomers. This study analyzed how real‑time molecular interaction analysis may be utilized in combination with perturbations in order to understand the kinetics of a ligand‑receptor interaction, as well as some of its associated intracellular processes. Our multiple‑temperature and ‑inhibitor assay setup renders it possible to follow the EGFR monomer, dimer and internalized populations in a detailed manner, allowing for a new perspective of the EGFR biology.
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March-April 2013
Volume 1 Issue 2

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Spandidos Publications style
Björkelund H, Gedda L, Malmqvist M and Andersson K: Resolving the EGF-EGFR interaction characteristics through a multiple-temperature, multiple-inhibitor, real-time interaction analysis approach. Mol Clin Oncol 1: 343-352, 2013
APA
Björkelund, H., Gedda, L., Malmqvist, M., & Andersson, K. (2013). Resolving the EGF-EGFR interaction characteristics through a multiple-temperature, multiple-inhibitor, real-time interaction analysis approach. Molecular and Clinical Oncology, 1, 343-352. https://doi.org/10.3892/mco.2012.37
MLA
Björkelund, H., Gedda, L., Malmqvist, M., Andersson, K."Resolving the EGF-EGFR interaction characteristics through a multiple-temperature, multiple-inhibitor, real-time interaction analysis approach". Molecular and Clinical Oncology 1.2 (2013): 343-352.
Chicago
Björkelund, H., Gedda, L., Malmqvist, M., Andersson, K."Resolving the EGF-EGFR interaction characteristics through a multiple-temperature, multiple-inhibitor, real-time interaction analysis approach". Molecular and Clinical Oncology 1, no. 2 (2013): 343-352. https://doi.org/10.3892/mco.2012.37