A semisynthetic protease DNA-nanomachine

• Applicant: Professorin Dr. Barbara Saccà
• Subject Area: Biological and Biomimetic Chemistry
• Term: from 2017 to 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 388607897

The main goal of this project is to generate a semisynthetic protease DNA-nanomachine, i.e. a compartmentalized multi-catalytic system capable of performing regulated proteolysis. We will employ DNA origami methodologies to create tubular shaped compartments, which will be individually loaded in their inner cavity with a distinct number, type and geometric arrangement of proteases. The DNA-encaged enzymes will be then combined in a lane through stepwise assembly, with variable stoichiometry and/or sequential order of the single chambers. The accessibility of unfolded polypeptides to the catalytic core of the degradation machine will be regulated by pores of variable sizes at both entries of the channel and/or removal of the terminal caps. Thus, emulating the building principles of the proteasome, one of the most fascinating proteolytic machines in nature, we aim at elucidating the impact of spatially controlled compartmentalization on the performance of proteolytic cascades. We envisage that the modularity of our design and its applicability to other enzyme classes will enable the generation of multifunctional biohybrid compartments, whose programmable structural and functional complexity may help to provide fundamental understanding of the role of spatial constraint and serial processing in the regulation of enzymatic reactions.

• DFG Programme: Research Grants
• International Connection: Denmark
• Cooperation Partner: Professorin Victoria Birkedal