RRC ID 67017
Author Pascarelli S, Merzhakupova D, Uechi GI, Laurino P.
Title Binding of single-mutant epidermal growth factor (EGF) ligands alters the stability of the EGF receptor dimer and promotes growth signaling.
Journal J Biol Chem
Abstract The epidermal growth factor receptor (EGFR) is a membrane-anchored tyrosine kinase that is able to selectively respond to multiple extracellular stimuli. Previous studies have indicated that the modularity of this system may be caused by ligand-induced differences in the stability of the receptor dimer. However, this hypothesis has not been explored using single-mutant ligands thus far. Herein, we developed a new approach to identify residues responsible for functional divergence by selecting residues in the epidermal growth factor (EGF) ligand that are conserved among orthologs yet divergent between paralogs. Then, we mutated these residues and assessed the mutants' effects on the receptor using a combination of molecular dynamics (MD) and biochemical techniques. Although the EGF mutants had binding affinities for the EGFR comparable with the WT ligand, the EGF mutants showed differential patterns of receptor phosphorylation and cell growth in multiple cell lines. The MD simulations of the EGF mutants indicated that mutations had long-range effects on the receptor dimer interface. This study shows for the first time that a single mutation in the EGF is sufficient to alter the activation of the EGFR signaling pathway at the cellular level. These results also support that biased ligand-receptor signaling in the tyrosine kinase receptor system can lead to differential downstream outcomes and demonstrate a promising new method to study ligand-receptor interactions.
Volume 297(1)
Pages 100872
Published 2021-7-1
DOI 10.1016/j.jbc.2021.100872
PII S0021-9258(21)00672-4
PMID 34126069
PMC PMC8259408
MeSH 3T3 Cells Animals Binding Sites Cell Line, Tumor Cell Proliferation Epidermal Growth Factor / genetics* ErbB Receptors / metabolism* Humans Mice Molecular Dynamics Simulation Mutation* Protein Binding Protein Stability Signal Transduction*
IF 4.238
Human and Animal Cells 3T3 Swiss Albino(RCB1642) A431(RCB0202)