RT Journal Article
SR Electronic
T1 The efficacy of EphA2 tyrosine phosphorylation increases with EphA2 oligomer size
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.06.07.495003
DO 10.1101/2022.06.07.495003
A1 Zapata-Mercado, Elmer
A1 Biener, Gabriel
A1 McKenzie, Daniel
A1 Wimley, William C.
A1 Pasquale, Elena B.
A1 Raicu, Valerica
A1 Hristova, Kalina
YR 2022
UL http://biorxiv.org/content/early/2022/06/09/2022.06.07.495003.abstract
AB The receptor tyrosine kinase (RTK) EphA2 is expressed in epithelial and endothelial cells and controls the assembly of cell-cell junctions. EphA2 has also been implicated in many diseases, including cancer. Unlike most RTKs, which signal predominantly as dimers, EphA2 readily forms higher order oligomers upon ligand binding. Here we investigated if a correlation exists between EphA2 signaling properties and the size of the EphA2 oligomers induced by multiple ligands, including the widely used ephrinA1-Fc ligand, the soluble monomeric m-ephrinA1, and novel engineered peptide ligands. We used Fluorescence Intensity Fluctuation (FIF) spectrometry to characterize the EphA2 oligomer populations induced by the different ligands. Interestingly, we found that different monomeric and dimeric ligands induce EphA2 oligomers with widely different size distributions. Comparison of FIF brightness distribution parameters and EphA2 signaling parameters reveals that the efficacy of EphA2 phosphorylation on tyrosine 588, which is indicative of receptor activation, correlates with EphA2 mean oligomer size. However, other characteristics, such as the efficacy of AKT inhibition and ligand bias coefficients, appear to be independent of EphA2 oligomer size. This work highlights the utility of FIF in RTK signaling research and demonstrates a quantitative correlation between the architecture of EphA2 signaling complexes and signaling features.Competing Interest StatementThe authors have declared no competing interest.