Understanding Evolution of Protein Function Through Design

The work proposed in Project 1 is based on the observation that one can design short Catalytic AMyloid- forming Peptides (CAMPs) to catalyze chemical reactions with high efficiency in addition to their own self- assembly. The goals of the proposed work are: 1. to understand the structural and mechanistic basis for the very high activity of CAMPs; 2. to achieve improved catalytic efficiency and substrate selectivity in CAMPs; 3. to create amyloid-organic frameworks, a new class of catalytic materials; 4. to develop complex multifunctional, light-driven and regulated CAMPs with tunable properties for synthesis of complex products. Development and characterization of catalytic amyloids will advance several fields of biomedical importance. It will set important structural and functional reference points for the broad community of scientists interested in the role of amyloids in protein folding, catalysis and health The structure-activity relationships and structural insights generated in the proposed work will help us better understand the mechanisms of amyloid toxicity and will improve our knowledge of the structures adopted by more complex amyloid-forming proteins. In addition to its practical value, this research program will have a profound impact on our understanding of the fundamental aspects of catalysis. Project 2 aims to develop a new NMR-based experimental approach to guide directed evolution to fully realize its potential in repurposing enzymes for new functions. Specifically we will: 1. gain a thorough understanding of the limits and the applicability of the method; 2. Independently validate the approach in different protein scaffolds; 3. use NMR guided directed evolution to create high efficient and selective catalysts for practically important chemical transformations. Project 3. The ability of pathogens to neutralize drugs via a newly developed catalytic activity is one of the mechanisms of drug resistance. Therefore, a deeper understanding of the factors that determine the ability of proteins to catalyze new chemical transformations is of paramount importance. We aim to determine the factors that guide evolution of protein function at a molecular level and use these principles to create catalysts for chemical transformations not found in nature. We will combine a minimalist computational approach with sophisticated protein engineering tools to create new protein catalysts for a number of different chemical transformations including those that require metal cofactors.