Abstract
Despite its low cellular abundance, phosphotyrosine (pTyr) regulates numerous cell signaling pathways in health and disease. We applied comprehensive phosphoproteomics to unravel differential regulators of receptor tyrosine kinase (RTK)-initiated signaling networks upon activation by Pdgf-ββ, Fgf-2, or Igf-1 and identified more than 40,000 phosphorylation sites, including many phosphotyrosine sites without additional enrichment. The analysis revealed RTK-specific regulation of hundreds of pTyr sites on key signaling molecules. We found the tyrosine phosphatase Shp-2 to be the master regulator of Pdgfr pTyr signaling. Application of a recently introduced allosteric Shp-2 inhibitor revealed global regulation of the Pdgf-dependent tyrosine phosphoproteome, which significantly impaired cell migration. In addition, we present a list of hundreds of Shp-2-dependent targets and putative substrates, including Rasa1 and Cortactin with increased pTyr and Gab1 and Erk1/2 with decreased pTyr. Our study demonstrates that large-scale quantitative phosphoproteomics can precisely dissect tightly regulated kinase-phosphatase signaling networks.
| Original language | English |
|---|---|
| Journal | Cell Reports |
| Volume | 22 |
| Issue number | 10 |
| Pages (from-to) | 2784-2796 |
| Number of pages | 14 |
| ISSN | 2211-1247 |
| DOIs | |
| Publication status | Published - 2018 |
Keywords
- label-free quantitation
- mass spectrometry
- orbitrap
- PDGF
- phosphoproteomics
- Q exactive
- Shp-2
- SHP099
- TiO2
- tyrosine phosphorylation