Artificial Enzyme Design and Assembly
The design of new proteins and enzymes remains one of the great challenges in biochemistry and tests our fundamental understanding of the nature of protein as a material. Unlocking the exceptionally diverse and powerful array of chemistries that natural enzymes perform promises routes to new drugs, therapies and green industrial processes.
Most approaches to this end have focused on modifying natural proteins and enzymes to impart new or altered catalytic function with varying degrees of success. The problems associated with such alterations are due to the layers of complexity that nature incorporates through natural selection into a protein’s complicated three-dimensional structure.
Simplified synthetic protein scaffolds offer a means to avoid such complexity, learn the principles guiding functional protein assembly and render the modular assembly of enzymatic function a tangible reality. This approach is illustrated through the assembly of an artificial O2 binding protein (Koder and Anderson, Nature, 2009) that reproduces the function of natural proteins such as neuroglobin and myoglobin in a simple heme-binding 4-helix bundle untouched by natural selection. The tractable design process resolves the roles of individual amino acids with their function and opens the door to the powerful oxygenic catalysis common to heme-containing dioxygenases and cytochromes P450.
We aim to use this approach for the construction of artificial protein modules that, when assembled, integrate functional elements common to natural redox enzymes (electron/proton transfer, ligand/substrate binding and light harvesting) to create artificial oxidoreductases.
Watkins DW, Armstrong CT, Anderson JLR. (2014) De novo protein components for oxidoreductase assembly and integration. Current Opinion in Chemical Biology. In press (DOI:10.1016/j.cbpa.2014.01.016)
Anderson JLR, Armstrong CT, Kodali G, Lichtenstein BR, Watkins DW, Mancini JA, Boyle AL, Farid TA, Crump MP, Moser CC, Dutton PL. (2014) Constructing a man-made c-type cytochrome maquette in vivo: electron transfer, oxygen transport and conversion to a photoactive light harvesting maquette. Chemical Science. 5(2):507-514
Farid TA, Kodali G, Solomon LA, Lichtenstein BR, Sheehan MM, Fry BA, Bialas C, Ennist NM, Siedlecki JA, Zhao ZY, Stetz MA, Valentine KG, Anderson JLR, Wand AJ, Discher BM, Moser CC, Dutton PL. (2013) Elementary tetrahelical protein design for diverse oxidoreductase functions. Nature Chemical Biology. 9(12):826-833
Armstrong CT, Watkins DW, Anderson JLR. (2013) Constructing manmade enzymes for oxygen activation. Dalton Trans. 42(9):3136-3150