cAMP signalling -novel downstream targets of PKA and Epac identified by phospho-proteomics.

I. Implement novel method for quantitative COFRADIC non-gel global phospho-proteomics through stay of candidate in the laboratory of prof. J. Vandenkerckhove at Univ. of Gent II. The phospho-proteome will be compared in cells in response to activation of PKA (by N6-MB-cAMP) or Epac (by 8-pCPT-2´-O-Me-cAMP) III. The proteomics data will be compared to single cell flow-cytometry using selected phospho-epitope specific antibodies. This will allow estimation of cell-to-cell variation and cell cycle dependence of selected phosphorylation events IV. Based on II, III candidate phosphoproteins will be sought identified. V. Comparative holoenzyme formation studies will also be performed with the pure proteins. VI. Develop improved cAMP analog agonists and anta gonists discriminating isozyme I and II of PKA