Signalling Cascades, Protein Quantification and Reverse Phase Protein Microarrays

Robert Wild

The quantitative monitoring of proteins and their modifications can serve as a measurement for their activity. Coordinated activity within a widespread network of intracellular signalling pathways is needed in order to process cellular information. Numerous intracellular signalling cascades have been analysed and documented. Lots of information lies within the kinetics and the interaction between different pathways. The project focuses on the attempt to reveal the underlying mechanisms of drug action in nociceptive neurons. The investigation and understanding of signalling cascades in nociceptive neurons could provide knowledge about the mode of action of drug targets used in pain therapy and treatment. Protein microarrays can be used to follow signalling cascades, post translational modifications and protein abundance. Reverse phase protein microarrays (RPPM) offer the potential to analyze small quantity samples, at several time points, with different sets of experiments and additionally several replicates at once. This technique will provide datasets allowing relative protein quantification and their role and state in signalling cascades. Furthermore information, that will give the opportunity to experimentally verify or contradict theoretical protein network models, can be obtained.

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