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Spatio-temporal regulation of growth factor signaling by PTPs in living cells

Final Report Summary - PTPS (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.