Discovery of Proteinase Inhibitors via High-Throughput Screening

Proteinases are enzymes that hydrolyze other proteins.  This class of enzymes has members that play key roles in metabolism and storage, regulation, signaling, and cell-cycle control.  A wide variety of microorganisms also requires the action of their own proteinases for normal function.  A widely known example is the proteinase of HIV, the inhibition of which controls the development of AIDS in human patients.  In addition to HIV, a number of other medically important viruses also have obligate proteinases, including the hepatitis C, West Nile, and Dengue viruses.  Likewise, most pathogenic bacteria have a periplasmic proteinase whose activity is required for infection and virulence.  Each of these proteinases represents attractive drug targets.  In order to streamline the development of new inhibitors of different classes of proteinases, we have created a semi-synthetic genetic switch for use with combinatorial genetics and high-throughput selection/screening to look at billions of different proteinase-inhibitor combinations simultaneously.  In addition to developing new inhibitors, the vast amount of data that can be collected for a particular ligand-receptor pair will allow us to evaluate structure-function hypotheses in the search for advanced therapeutic strategies.