Single-molecule imaging of EGFR signalling on the surface of living cells

  title={Single-molecule imaging of EGFR signalling on the surface of living cells},
  author={Yasushi Sako and Shigeru Minoghchi and Toshio Yanagida},
  journal={Nature Cell Biology},
The early events in signal transduction from the epidermal growth factor (EGF) receptor (EGFR) are dimerization and autophosphorylation of the receptor, induced by binding of EGF. Here we observe these events in living cells by visualizing single molecules of fluorescent-dye-labelled EGF in the plasma membrane of A431 carcinoma cells. Single-molecule tracking reveals that the predominant mechanism of dimerization involves the formation of a cell-surface complex of one EGF molecule and an EGFR… 

Single‐molecule analysis of epidermal growth factor binding on the surface of living cells

A small number of EGFR on the cell surface formed dimericbinding sites, which bound EGF two orders of magnitude faster than the monomeric binding sites, and these two mechanisms facilitate the formation of signaling dimers of EGF/EGFR complexes.

Reversible dimerization of EGFR revealed by single-molecule fluorescence imaging using quantum dots.

Intermolecular interactions involved in the lateral propagation of cell-signaling by epidermal growth factor receptors (EGFRs) are explored and it is found that signaling dimers of EGFR are continuously formed in cell membrane through reversible association of heterodimers [EGF(EGFR)(2].

Optical bioimaging: from living tissue to a single molecule: single-molecule visualization of cell signaling processes of epidermal growth factor receptor.

This work has studied the process of early signal transduction of epidermal growth factor in single molecules, and suggested that this highly sensitive response to EGF was due to the amplification of the EGFR signal using dynamic clustering, reorganization of the dimers, and lateral mobility of EGFR on the cell surface.

Single-Molecule Imaging of Signaling Molecules in Living Cells

Using the single-molecule technique, the process of dimerization, a key step of EGFR signal transduction was observed and detection of single molecules was confirmed by single-step photobleaching.

Single-molecule fluorescence detection of the epidermal growth factor receptor (EGFR) in membrane discs

This work reports the development and characterization of functional fluorescently-labeled full-length EGFR in model membrane nanolipoprotein particles (NLPs) for in vitro fluorescence studies and investigates ATP-EGFR interactions.

Single-molecule imaging reveals the stoichiometry change of epidermal growth factor receptor during transactivation by β2-adrenergic receptor

It is found that although EGFR existed as monomers at the plasma membrane of resting cells, they became dimers and thus diffused slower following the activation of β2-adrenergic receptor (β2-AR) by isoproterenol (ISO), and it is proved that β-AR-mediated changes of EGFR in stoichiometry and dynamics were mediated by Src kinase.

Evidence for extended YFP-EGFR dimers in the absence of ligand on the surface of living cells

The results suggest that EGF-induced activation occurs within or between pre-formed and extended dimers with very little change in the extension of the N-terminii from the membrane surface.

EGF signalling amplification induced by dynamic clustering of EGFR.

Single molecule imaging and FLIM show different structures for high and low-affinity EGFRs in A 431 cells

The results support a structural model in which asymmetric EGFR head-to-head interfaces may be relevant for HA EGFR oligomerisation, and measure the inter-EGF distances within discrete EGF pairs and the vertical distance from EGF to the plasma membrane.



Two EGF molecules contribute additively to stabilization of the EGFR dimer

The proposed model of EGF‐induced sEGFR dimerization suggests possible mechanisms for both ligand‐induced homo‐ and heterodimerization of the EGFR (or erbB) family of receptors.

Self-phosphorylation of epidermal growth factor receptor: evidence for a model of intermolecular allosteric activation.

An allosteric aggregation model is formulated for the activation of the cytoplasmic kinase function of the receptor by EGF because this model may be relevant to the mechanism by which the mitogenic signal of EGF is transferred across the membrane.

Heterogeneity of epidermal growth factor binding kinetics on individual cells.

Immunodetection of the ligand-activated receptor for epidermal growth factor.

A method to detect the activation state of Epidermal Growth Factor receptor (EGFr) with a monoclonal antibody (mAb74), found to preferentially recognize the ligand-activated EGFr as detected by immunoprecipitation, Western blotting and immunocytochemical techniques.

Epidermal growth factor induces rapid, reversible aggregation of the purified epidermal growth factor receptor.

It is concluded that receptor oligomerization is an intrinsic property of the occupied EGF receptor and that it may play a role in the activation of the kinase function and the subsequent transmembrane signaling process.

Epidermal growth factor receptors associated to cytoskeletal elements of epidermoid carcinoma (A431) cells

Using stereo micrographs of replica's obtained from isolated cytoskeletons, it is shown that gold-labeled EGF receptors are exclusively present on the cortical membrane-associated region of the cytoskeleton and not on more intracellular-located filaments.