Catalysis and Electron Transfer in De Novo Designed Helical Scaffolds

: The relationship between protein structure and function is one of the greatest puzzles within biochemistry. De novo metalloprotein design is a way to wipe the board clean and determine what is required to build in function from the ground up in an unrelated structure. This review focuses on protein design efforts to create de novo metalloproteins within alpha helical scaffolds. Examples  of successful designs from our lab include those with carbonic anhydrase or nitrite reductase activity by incorporating a ZnHis3 or CuHis3 site, or that recapitulate the spectroscopy of unique electron transfer sites from cupredoxins (CuHis2Cys) to rubredoxin (FeCys4). This work showcases the versatility of alpha helices as scaffolds for metalloprotein design and the progress that is possible through careful rational design as we show the invariance of carbonic anhydrase activity to site position and scaffold, refine our cupredoxin models, and enhance nitrite reductase activity up to 1000-fold.