Related Experiment Video

Updated: Mar 16, 2026

Live-Cell Förster Resonance Energy Transfer Imaging of Metabolically Regulated Akt Activation Dynamics in HepG2 Cells
08:03

Live-Cell Förster Resonance Energy Transfer Imaging of Metabolically Regulated Akt Activation Dynamics in HepG2 Cells

Published on: May 23, 2025

861

Similar requirement for clathrin in EGF- and HGF- stimulated Akt phosphorylation

Stefanie Lucarelli1, Rohan Pandey2, Gurjeet Judge1

  • 1Department of Chemistry and Biology, Ryerson University, Toronto, Ontario, Canada; Graduate Program in Molecular Science, Ryerson University, Toronto, Ontario, Canada.

Communicative & Integrative Biology
|August 5, 2016
PubMed

Insights

Clathrin regulates signaling pathways for epidermal growth factor (EGF) receptor and Met. This study shows clathrin is crucial for Met-induced Akt phosphorylation, similar to its role in EGF receptor signaling.

Area of Science:

  • Cell Biology
  • Molecular Signaling

Background:

  • Receptor tyrosine kinases like EGFR and Met activate intracellular signals, including Akt, which controls cell survival and proliferation.
  • Receptor activation triggers recruitment into clathrin-coated pits (CCPs) and subsequent endocytosis.

Purpose of the Study:

  • To investigate if clathrin regulates Akt signaling downstream of Met, similar to its established role in EGFR signaling.
  • To determine the role of clathrin in Met-mediated signaling pathways.

Main Methods:

  • Stimulation of ARPE-19 cells with Hepatocyte Growth Factor (HGF), the Met ligand.
  • Utilizing the clathrin inhibitor pitstop2 to perturb clathrin function.
  • Analyzing the enrichment of phosphorylated Gab1 (pGab1) within CCPs.
  • Measuring Akt phosphorylation levels.

Main Results:

  • HGF stimulation resulted in the enrichment of pGab1 within CCPs in ARPE-19 cells.
  • Inhibition of clathrin with pitstop2 significantly decreased HGF-stimulated Akt phosphorylation.
  • These findings suggest clathrin's involvement in Met signaling.

Conclusions:

  • Clathrin plays a regulatory role in Met signaling, leading to Akt phosphorylation.
  • The mechanism of clathrin's regulation of Met signaling appears similar to its role in EGFR signaling.
  • Clathrin-mediated regulation of Akt phosphorylation is a conserved mechanism across different receptor tyrosine kinases.
Keywords:
TIRF microscopyclathrinendocytosismembrane trafficphosphorylationreceptor tyrosine kinasesignaling adaptorsignaling microdomainsignaling scaffold

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