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Updated: May 11, 2026

Oligopeptide Competition Assay for Phosphorylation Site Determination
Published on: May 18, 2017
Novel binding partners and differentially regulated phosphorylation sites clarify Eps8 as a multi-functional adaptor
Debbie L Cunningham1, Andrew J Creese, Giulio Auciello
1Cancer Research UK Growth Factor Signalling Group, School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Birmingham, United Kingdom.
This study identifies new phosphorylation sites on Eps8 protein, revealing its regulation by fibroblast growth factor receptor (FGFR) and Src kinases. It also uncovers novel binding partners involved in cell signaling and vesicle trafficking.
Area of Science:
- Cell Biology
- Molecular Biology
- Proteomics
Background:
- Eps8 protein participates in cell signaling and receptor trafficking.
- Eps8 is phosphorylated by fibroblast growth factor receptor (FGFR) and Src kinases.
- Understanding Eps8 phosphorylation is crucial for receptor signaling and trafficking pathways.
Purpose of the Study:
- To identify specific FGFR and Src family kinase-dependent phosphosites on Eps8.
- To discover novel phosphodependent binding partners of Eps8.
- To elucidate Eps8's role at the intersection of receptor signaling and vesicular trafficking.
Main Methods:
- Differential proteomic analysis of Eps8.
- Identification of phosphorylation sites (pTyr and pSer/Thr).
- Peptide affinity purification to identify binding partners.
Main Results:
- 22 Eps8 phosphorylation sites identified, including Src and FGFR kinase-dependent sites.
- Novel Eps8 binding partners discovered, including clathrin, AP-2, NBR1, Vav2, IRS4, and Shp2.
- Confirmed Eps8's involvement in regulating receptor trafficking and signaling.
Conclusions:
- This study expands the understanding of Eps8 post-translational modifications.
- Identified novel Eps8 binding partners crucial for receptor trafficking and signaling.
- Confirmed Eps8's function linking receptor signaling and vesicular transport.
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