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Updated: Jun 15, 2026

Identification of Kinase-substrate Pairs Using High Throughput Screening
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Differential phosphoproteomics of fibroblast growth factor signaling: identification of Src family kinase-mediated

Debbie L Cunningham1, Steve M M Sweet, Helen J Cooper

  • 1School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom.

Journal of Proteome Research
|March 16, 2010
PubMed

Insights

This study identifies 80 Src family kinase-dependent phosphorylation events and 18 Src family kinase-dependent Dok1 interactions in fibroblast growth factor signaling. Six novel Src family kinase-dependent phosphorylation sites on Dok1 were discovered.

Area of Science:

  • Cellular signaling pathways
  • Receptor tyrosine kinase signaling
  • Protein phosphorylation

Background:

  • Fibroblast growth factor receptor (FGFR) activation initiates signal transduction via tyrosine phosphorylation.
  • Src family kinases (SFKs) play crucial roles in initiating and terminating FGF signaling cascades.
  • Understanding SFK-dependent phosphorylation is key to deciphering FGF signaling.

Purpose of the Study:

  • To identify SFK-dependent tyrosine phosphorylation events during FGF signaling.
  • To investigate SFK-dependent interactions and phosphorylation of the Dok1 adaptor protein.
  • To elucidate the role of SFKs in FGF-mediated cellular responses.

Main Methods:

  • Stable Isotope Labeling by Amino acids in Cell culture (SILAC) combined with chemical SFK inhibition.
  • High-coverage phosphopeptide mapping of the Dok1 protein.
  • Identification of SFK-dependent protein-protein interactions.

Main Results:

  • Identified 80 SFK-dependent phosphorylation events on 40 proteins in FGF-stimulated cells.
  • Discovered 18 SFK-dependent Dok1 interactions.
  • Mapped 9 SFK-dependent Dok1 phosphorylation sites, including 6 previously unknown sites.

Conclusions:

  • SFKs regulate a significant number of phosphorylation events and interactions in FGF signaling.
  • Dok1 is a key substrate and interaction partner regulated by SFKs in this pathway.
  • These findings provide novel insights into the molecular mechanisms of FGF signaling.

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