PTPS · Spatio-temporal regulation of growth factor signaling by PTPs in living cells
FP7 — People (Marie Curie Actions)
- Duration
- 2008-03-01 → 2010-02-28
- EU contribution
- €219,682
- Participants
- 1
- Scheme
- MC-IEF
Lines connect the coordinator with its partners.
Results in brief
Spatio-temporal regulation of growth factor signaling by PTPs in living cells
Quantification of posttranslational modifications in situ constitutes a crucial step in order to understand how signalling networks relay extracellular signals inside the cell and alter the cell's phenotype. We have developed a method for quantifying tyrosine phosphorylation in situ by using fluorescence lifetime imaging microscopy (FLIM) on cell arrays (CA-FLIM). CA-FLIM allows identifying tyrosine kinase/phosphatase substrates and quantifying phospho-tyrosine patterns in large networks, providing information on their structure and dynamics, also in spatially regulated cellular processes. Currently we are using CA-FLIM to systematically screen the PTP-ome for components of the signalling networks that act downstream EGFR activation. Finally, the molecular interactions and dynamic properties of protein tyrosine phosphatases are being studied by micro-spectroscopy techniques. This will provide relevant information about the subcellular partitioning of PTP activities inside the cell. The following points are treated: - Structure and dynamics of phospho-tyrosine signalling networks - Regulation of EGFR phosphorylation by protein tyrosine phosphatises.
Data: CORDIS, © European Union
Project objective
Receptor tyrosine kinases (RTK) are involved in the regulation of cell proliferation, survival, differentiation and motility upon binding of growth factors. Disturbance of the normal balance between tyrosine kinase and Protein Tyrosine Phosphatase (PTP) activity results in aberrant tyrosine phosphorylation and has been implicated in the etiology of several human diseases, including cancer, diabetes and inflammation. Thus, it is important to understand how phosphorylation is regulated by members of both enzyme families. The objective of this proposal is to analyze the spatial regulation of Epidermal Growth Factor (EGF) signaling by PTPs in intact living cells. For this we propose to use quantitative fluorescence lifetime imaging microscopy (FLIM) in combination with functional genomic approaches. We will obtain information about which PTPs regulate the kinase activity of the EGF receptor, and the biochemical conectivity among PTP activities in response to EGF and other growth factors or hormones. Finally we will also investigate how PTP activities are regulated in response to EGF and where this happens in the cell. This spatio-temporal regulation of PTP activities will be studied by a novel approach in which enzyme-substrate complexes are directly imaged in living cells. Observation of `live´ biochemistry on a microscopic level has the advantage of preserving the network interconnectivity and spatial organization, allowing the investigation of the molecular dynamics that gives rise to the specificity in intracellular signal transduction. Since cellular responses are generated by the action of different gene products, cellular biology demands the regulation of signaling pathways at a proteomic scale to be studied to understand how signal specificity is achieved in cells and how stimuli are integrated to develop specific responses. Thus, our goal will be to obtain new information about dynamic properties of signaling networks at the proteomic scale.
Original text from CORDIS.
Participants
- MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV · MUNCHENCoordinatorGermany
Links
Data: CORDIS, © European Union