The first sub-project involves the establishment of a platform for the generation and characterization of monobodies against specific target proteins such as the proto-oncogene Src family kinases involved in various cancers and the centriolar SAS-6 proteins necessary for centriole duplication and function. Further mechanistic studies are pursued to characterise in depth some of the mono-bodies studied. At a larger scope, the establishment of a platform for the generation of monobodies is extremely valuable for various projects in the NCCR.
Sub-project 2 involves the development of strategies to switch on- off- such monobodies and make these tools even more valuable.
In the third sub-project, expressed protein ligation methods are refined to produce proteins in a traceless manner with defined chemical modifications. These methods are used e.g. to generate α and β-tubulin proteins with defined post-translational modifications (PTMs) at their C-terminal tail and to characterize the importance of these modifications in asymmetric vesicle trafficking and in centriole formation.
Finally, the fourth sub-project involves the generation of new semi-synthetic fluorescent sensor proteins to measure with unprecedented precision and spatio-temporal resolution NADPH/NADP+ ratios or NAD+ concentrations within the cytosol or lumen of organelles in living cells. Next, general strategies for improving the dynamic range of such sensors are under continuous development.