Proteomic analyses of signalling complexes associated with receptor tyrosine kinase identify novel members of fibroblast growth factor receptor 3 interactome
Language English Country Great Britain, England Media print-electronic
Document type Journal Article
Grant support
R01 AR066124
NIAMS NIH HHS - United States
R01 DE019567
NIDCR NIH HHS - United States
PubMed
29030113
DOI
10.1016/j.cellsig.2017.10.003
PII: S0898-6568(17)30271-1
Knihovny.cz E-resources
- Keywords
- FGFR3, Fibroblast growth factor, interactome, receptor tyrosine kinase, signal transduction,
- MeSH
- Adaptor Proteins, Signal Transducing genetics metabolism MeSH
- NIH 3T3 Cells MeSH
- Cyclin-Dependent Kinases genetics metabolism MeSH
- Peptide Elongation Factor 1 genetics metabolism MeSH
- Endosomal Sorting Complexes Required for Transport genetics metabolism MeSH
- Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases genetics metabolism MeSH
- Phosphoproteins genetics metabolism MeSH
- Phosphorylation MeSH
- HEK293 Cells MeSH
- Homeodomain Proteins genetics metabolism MeSH
- Cyclin-Dependent Kinase-Activating Kinase MeSH
- Humans MeSH
- Protein Interaction Mapping MeSH
- Mice MeSH
- Protein Serine-Threonine Kinases genetics metabolism MeSH
- Proteomics methods MeSH
- Receptor, Fibroblast Growth Factor, Type 3 genetics metabolism MeSH
- Gene Expression Regulation * MeSH
- Signal Transduction genetics MeSH
- Gene Expression Profiling MeSH
- Protein Binding MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Adaptor Proteins, Signal Transducing MeSH
- CILK1 protein, human MeSH Browser
- Cyclin-Dependent Kinases MeSH
- EEF1A1 protein, human MeSH Browser
- Peptide Elongation Factor 1 MeSH
- Endosomal Sorting Complexes Required for Transport MeSH
- FGFR3 protein, human MeSH Browser
- Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases MeSH
- Phosphoproteins MeSH
- Homeodomain Proteins MeSH
- INPPL1 protein, human MeSH Browser
- Cyclin-Dependent Kinase-Activating Kinase MeSH
- MAK protein, human MeSH Browser
- Protein Serine-Threonine Kinases MeSH
- Receptor, Fibroblast Growth Factor, Type 3 MeSH
- SHOX2 protein, human MeSH Browser
- STAM protein, human MeSH Browser
Receptor tyrosine kinases (RTKs) form multiprotein complexes that initiate and propagate intracellular signals and determine the RTK-specific signalling patterns. Unravelling the full complexity of protein interactions within the RTK-associated complexes is essential for understanding of RTK functions, yet it remains an understudied area of cell biology. We describe a comprehensive approach to characterize RTK interactome. A single tag immunoprecipitation and phosphotyrosine protein isolation followed by mass-spectrometry was used to identify proteins interacting with fibroblast growth factor receptor 3 (FGFR3). A total of 32 experiments were carried out in two different cell types and identified 66 proteins out of which only 20 (30.3%) proteins were already known FGFR interactors. Using co-immunoprecipitations, we validated FGFR3 interaction with adapter protein STAM1, transcriptional regulator SHOX2, translation elongation factor eEF1A1, serine/threonine kinases ICK, MAK and CCRK, and inositol phosphatase SHIP2. We show that unappreciated signalling mediators exist for well-studied RTKs, such as FGFR3, and may be identified via proteomic approaches described here. These approaches are easily adaptable to other RTKs, enabling identification of novel signalling mediators for majority of the known human RTKs.
Department of Biology Faculty of Medicine Masaryk University 62500 Brno Czech Republic
Institute of Experimental Biology Faculty of Science Masaryk University 62500 Brno Czech Republic
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