Magnitude of Ubiquitination Determines the Fate of Epidermal Growth Factor Receptor Upon Ligand Stimulation.

Vyacheslav Akimov, Mirjam Fehling-Kaschek, Inigo Barrio-Hernandez, Michele Puglia, Jakob Bunkenborg, Mogens M Nielsen, Jens Timmer, Jörn Dengjel, Blagoy Blagoev

Journal: Journal of molecular biology 2021;433(21):167240

PMID: 34508725

Free Full Text on Elsevier Science View on PubMed

Abstract

Receptor tyrosine kinases (RTK) bind growth factors and are critical for cell proliferation and differentiation. Their dysregulation leads to a loss of growth control, often resulting in cancer. Epidermal growth factor receptor (EGFR) is the prototypic RTK and can bind several ligands exhibiting distinct mitogenic potentials. Whereas the phosphorylation on individual EGFR sites and their roles for downstream signaling have been extensively studied, less is known about ligand-specific ubiquitination events on EGFR, which are crucial for signal attenuation and termination. We used a proteomics-based workflow for absolute quantitation combined with mathematical modeling to unveil potentially decisive ubiquitination events on EGFR from the first 30 seconds to 15 minutes of stimulation. Four ligands were used for stimulation: epidermal growth factor (EGF), heparin-binding-EGF like growth factor, transforming growth factor-α and epiregulin. Whereas only little differences in the order of individual ubiquitination sites were observed, the overall amount of modified receptor differed depending on the used ligand, indicating that absolute magnitude of EGFR ubiquitination, and not distinctly regulated ubiquitination sites, is a major determinant for signal attenuation and the subsequent cellular outcomes.

Address: Center for Experimental BioInformatics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark.; Institut of Physics, University of Freiburg, Hermann-Herder-Str. 3, 79104 Freiburg, Germany.; Institut of Physics, University of Freiburg, Hermann-Herder-Str. 3, 79104 Freiburg, Germany; Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Schänzlestr. 18, 79104 Freiburg, Germany.; Department of Biology, University of Fribourg, Chemin du Musée 10, 1700 Fribourg, Switzerland. Electronic address: [email protected].; Center for Experimental BioInformatics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark. Electronic address: [email protected].

DOI: 10.1016/j.jmb.2021.167240

Link outs

Free resources

Full Text Sources:

https://linkinghub.elsevier.com/retrieve/pii/S0022...

Miscellaneous:

https://assays.cancer.gov/CPTAC-802

Research Materials:

https://antibodies.cancer.gov/detail/CPTC-EGFR-8

MeSH Terms: Amino Acid Sequence, Cell Line, Tumor, Epidermal Growth Factor, Epiregulin, Epithelial Cells, ErbB Receptors, Gene Expression, Heparin-binding EGF-like Growth Factor, Humans, Ligands, Models, Molecular, Mutation, Phosphorylation, Protein Conformation, Protein Processing, Post-Translational, Proteomics, Signal Transduction, Transforming Growth Factor alpha, Ubiquitination