A FRET-BASED SCREEN FOR THE BINDING OF COMPUTATIONALLY DESIGNED PROTEIN PAIRS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. One goal of computational biology is to have the ability to design novel protein partners. In this way novel protein activities could be created that may find use in both basic research, and in creating novel diagnostics or treatments for human disease. Now in an early phase of development, the process is to begin with two proteins that do not bind to each other, and to redesign them to create a specific, high affinity shared binding surface. We have begun by applying this methodology to design a pair of proteins, which interact via a novel protein-protein interface. Since this interface is solely the product of computational design, it is completely orthogonal to all proteins which comprise the yeast proteome. This feature suggests an interesting application as a set of affinity tags. A limiting step in the process is the assessment of whether changes have improved binding affinities. This project aims to combine the YRC expertise in both protein design and FRET to develop a method for rapid screening of the redesigned proteins for their ability to form heterodimers.