Project Grant R35GM155013

METALLOPROTEINS: EVOLUTION, CATALYSIS AND MATERIAL DESIGN - PROJECT 1: RECENTLY, WE HAVE DEMONSTRATED FOR THE FIRST TIME THE PREDICTIVE LINK BETWEEN NMR CHEMICAL SHIFT PERTURBATION, PROTEIN DYNAMICS AND PROBABILITY OF FINDING PRODUCTIVE MUTATIONS, WHICH ALLOWED US TO CREATE AN UNNATURAL METALLOENZYME WITH WITH A 107-FOLD IMPROVEMENT OVER THE BACKGROUND RATE AND LESS THAN 100-FOLD AWAY FROM THE DIFFUSION LIMIT. I PROPOSE TO EXPAND NMR-GUIDED DIRECTED EVOLUTION TO 1) UTILIZE OTHER PROTEIN DYNAMICS PROBING MODALITIES TO AAPLY THE APPROACH TO LARGER PROTEINS; 2) DEVELOP CATALYSTS FOR PRACTICALLY USEFUL REACTIONS; 3) DEVELOP OF METAL-BINDING PROTEINS WITH HIGH METAL AFFINITY. RESULTS GENERATED IN THE PROPOSED WORK WILL HELP ADVANCE FUNDAMENTAL UNDERSTANDING OF THE PRINCIPLES THAT DEFINE FUNCTION IN METALLOPROTEINS AND DRIVE ENZYMATIC ACTIVITY. FROM THE PRACTICAL STANDPOINT, THIS WORK WILL ALLOW FOR CREATION OF SMALLER FUNCTIONAL PROTEINS CAPABLE OF PROMOTING CHEMICAL TRANSFORMATIONS AND PRODUCE INEXPENSIVE AND BIOCOMPATIBLE PROTEIN CATALYSTS THAT COULD BE EASILY HANDLED IN THE APPLIED SETTING FOR PESTICIDE/CHEMICAL WEAPONS REMEDIATION, DNA HYDROLYSIS AND PHENOL OXIDATION. PROJECT 2: I WILL DEVELOP NEW APPROACHES TO OSTEOARTHRITIS TREATMENT USING SMART, STIMULI- RESPONSIVE, SELF-HEALING BIOCOMPATIBLE ANTIMICROBIAL HYDROGELS. IN ADDITION TO DIRECT BIOMEDICAL APPLICATION OF THE DESIGNED HYDROGEL MATERIALS, STRUCTURAL MODELS FOR THE HYDROGEL ASSEMBLIES WILL FACILITATE OUR UNDERSTANDING OF THE MECHANISM OF GELATION OF METALLOPEPTIDES AND WILL GUIDE RATIONAL DESIGN OF IMPROVED MATERIALS. 1