MilliporeSigma
  • Interdependence between EGFR and Phosphatases Spatially Established by Vesicular Dynamics Generates a Growth Factor Sensing and Responding Network.

Interdependence between EGFR and Phosphatases Spatially Established by Vesicular Dynamics Generates a Growth Factor Sensing and Responding Network.

Cell systems (2018-08-27)
Angel Stanoev, Amit Mhamane, Klaus C Schuermann, Hernán E Grecco, Wayne Stallaert, Martin Baumdick, Yannick Brüggemann, Maitreyi S Joshi, Pedro Roda-Navarro, Sven Fengler, Rabea Stockert, Lisaweta Roßmannek, Jutta Luig, Aneta Koseska, Philippe I H Bastiaens
ABSTRACT

The proto-oncogenic epidermal growth factor receptor (EGFR) is a tyrosine kinase whose sensitivity to growth factors and signal duration determines cellular behavior. We resolve how EGFR's response to epidermal growth factor (EGF) originates from dynamically established recursive interactions with spatially organized protein tyrosine phosphatases (PTPs). Reciprocal genetic PTP perturbations enabled identification of receptor-like PTPRG/J at the plasma membrane and ER-associated PTPN2 as the major EGFR dephosphorylating activities. Imaging spatial-temporal PTP reactivity revealed that vesicular trafficking establishes a spatially distributed negative feedback with PTPN2 that determines signal duration. On the other hand, single-cell dose-response analysis uncovered a reactive oxygen species-mediated toggle switch between autocatalytically activated monomeric EGFR and the tumor suppressor PTPRG that governs EGFR's sensitivity to EGF. Vesicular recycling of monomeric EGFR unifies the interactions with these PTPs on distinct membrane systems, dynamically generating a network architecture that can sense and respond to time-varying growth factor signals.

MATERIALS
Product Number
Brand
Product Description

Millipore
CellASIC ONIX switching plate mammalian cells (4 chamber), The M04 switching plates enable continuous perfusion culture for live cell analysis. Laminar flow provides rapid & uniform solution exchange for maximum cell health. The glass coverslide bottom surface ensures the highest resolution optical viewing.
Supelco
5,5-Dimethyl-1,3-cyclohexanedione, for HPLC derivatization, for the determination of aldehyde formaldehyde, ≥99.0%
Sigma-Aldrich
hEGF, EGF, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Poly-L-lysine hydrobromide, mol wt 70,000-150,000, lyophilized powder, γ-irradiated, BioXtra, suitable for cell culture
Sigma-Aldrich
DL-Cysteine, technical grade
Sigma-Aldrich
Anti-Sulfenic Acid Modified Cysteine (2-Thiodimedone-Specific Ig) Antibody, serum, from rabbit
Sigma-Aldrich
Cholera Toxin from Vibrio cholerae, ≥90% (SDS-PAGE), lyophilized powder