Quantifying the Interaction between EGFR Dimers and Grb2 in Live Cells

Nuala Del Piccolo, Kalina A Hristova

Research output: Contribution to journalArticle

Abstract

Adaptor proteins are a class of cytoplasmic proteins that bind to phosphorylated residues in receptor tyrosine kinases and trigger signaling cascades that control critically important cellular processes, such as cell survival, growth, differentiation, and motility. Here, we seek to characterize the interaction between epidermal growth factor receptor (EGFR) and the cytoplasmic adaptor protein growth factor receptor-bound protein 2 (Grb2) in a cellular context. To do so, we explore the utility of a highly biologically relevant model system, mammalian cells under reversible osmotic stress, and a recently introduced Förster resonance energy transfer microscopy method, fully quantified spectral imaging. We present a method that allows us to quantify the stoichiometry and the association constant of the EGFR-Grb2 binding interaction in the plasma membrane, in the presence and absence of activating ligand. The method that we introduce can have broad utility in membrane protein research, as it can be applied to different membrane protein-cytoplasmic protein pairs.

Original languageEnglish (US)
JournalBiophysical Journal
DOIs
StateAccepted/In press - 2017

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GRB2 Adaptor Protein
Epidermal Growth Factor Receptor
Membrane Proteins
Proteins
Energy Transfer
Osmotic Pressure
Receptor Protein-Tyrosine Kinases
Protein Binding
Microscopy
Cell Survival
Cell Membrane
Ligands
Growth
Research

ASJC Scopus subject areas

  • Biophysics

Cite this

Quantifying the Interaction between EGFR Dimers and Grb2 in Live Cells. / Del Piccolo, Nuala; Hristova, Kalina A.

In: Biophysical Journal, 2017.

Research output: Contribution to journalArticle

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