NON-TECHNICAL SUMMARY: The ability to precisely design and assemble three-dimensional biomaterials will open the door for significant impacts in many fields including biotechnology, bioremediation, drug delivery, and advanced medical technologies. To this end, this project will develop, validate, and apply novel approaches for the design and precise assembly of proteins, containing non-natural amino acids, into three-dimensional frameworks to create new classes of functional biomaterials. The proposed approach promises to be readily extensible to a variety of systems and applications, thereby making it an exciting new method for biomaterials design. In addition, this proposal includes research and teaching activities in biochemistry, structural biology, protein engineering, and chemical biology that provide unique opportunities for training highly interdisciplinary students and broadening participation in the future STEM workforce of the United States. TECHNICAL SUMMARY: The primary goal of the proposed research is to develop a general method for the assembly of large, ordered 3D protein biomaterials. Standard protein crystals are typified by properties including close packing between protein partners and a relative inability to design or control the lattices in which the assemblies form. The ability to control the geometric relationships and distances between assembled proteins within the lattice would allow for the development of a new class of ordered 3D protein m