A New Temperature-Dependent Strategy to Modulate the Epidermal Growth Factor Receptor

Biomaterials. 2018 Nov;183:319-330. doi: 10.1016/j.biomaterials.2018.07.063. Epub 2018 Aug 11.


The dynamic manipulation of kinases remains a major obstacle to unraveling cell-signaling networks responsible for the activation of biological systems. For example, epidermal growth factor (EGF) stimulates the epidermal growth factor receptor (EGFR/ErbB1); however, EGF also recruits other kinases (HER2/ErbB2) involved with various signaling pathways. To better study EGFR we report a new strategy to selectively activate receptor tyrosine kinases fused to elastin-like polypeptides (ELPs), which can be visualized inside mammalian cells using fixed and live-cell fluorescence microscopy. ELPs are high molecular weight polypeptides that phase separate abruptly upon heating. When an EGFR-ELP fusion is heated, it clusters, initiates receptor internalization, phosphorylates, initiates downstream kinase signaling, and undergoes retrograde transport towards the cell body. Unlike other strategies to block EGFR (small molecule inhibitors, RNAi, or transcriptional regulators), EGFR-ELP clustering can be specifically switched on or off within minutes. Live-cell imaging suggests that EGFR-ELPs assemble in most cells with only a 3 °C increase in temperature. This strategy was found reversible and able to dynamically control the downstream phosphorylation/activation of the ERK1/2 pathway. For the first time, this strategy enables the rational engineering of specific temperature-sensitive receptors that may have broad applications in the study and manipulation of biological processes.

Keywords: 293T; ERK activity dynamics; Elastin-like polypeptides; Green-fluorescent protein; Synthetic biology; Thermo-responsive protein-polymer.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Epidermal Growth Factor / chemistry
  • Epidermal Growth Factor / metabolism*
  • ErbB Receptors / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Microscopy, Fluorescence / methods
  • Peptides / genetics
  • Peptides / metabolism*
  • Phosphorylation
  • Protein-Tyrosine Kinases / genetics
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Temperature
  • Transfection


  • Peptides
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Epidermal Growth Factor
  • ErbB Receptors
  • Protein-Tyrosine Kinases