Team:Bonn

Making versatile control of biological machines easily possible for everybody we engineer a system for light dependend control of protein activity per protein degradation. For protein degradation we harness the ClpXP protease system of prokaryotes. This system enables to activate degradation of a peptid tag (ssrA) fused protein upon induction with the adaptor protein SspB. Using a Split version of SspB, protein degradation is activated through heterodimerisation of both SspB parts. As a proof of principle we investigate degradation of the fluorescent reporter protein mCherry. To obtain light inducible degradation we use a light dependend heterodimerisation system. The light sensitive part of this system is LOV2 (A. Sativa). Modelling of the light dependency of protein degradation will even make simple and accurate control of protein activity for any desired level of activity possible.

To enable protein degradation we decided on one proteases system (ClpXP), which allows specific degradation of proteins, is highly conserved and well established. For light dependency we compared several systems to finally find the best suited one. We created a comprehensive overview of all systems making artificially light control possible so that everyone can easily choose the optimal system for his approach.