Abstract 1921 Crystallization of a Diadenosine Polyphosphatase of the Nudix Hydrolase Superfamily from M. tuberculosis

M. tuberculosis contains 11 potential Nudix hydrolases, and we are characterizing these enzymes as potential novel antibiotic targets. The diadenosine polyphosphatases (ApnAases) / mRNA decapping enzymes are a family of enzymes within the Nudix hydrolase superfamily. In M. tuberculosis there is the primary Nudix ApnAase and the secondary Nudix ApnAase. The diadenosine polyphosphatases from Legionella pneumophilia and Bartonella bacilliformis have been found to be important in each pathogen's ability to invade its host cells. We are interested in whether these enzymes act in the same way in M. tuberculosis. If they are found to be involved in invasiveness and thus in virulence, then these enzymes could be novel antibiotic targets. Solving the X-ray crystal structure of this enzyme could be used for novel drug design against M. tuberculosis. We have cloned and overexpressed the primary Nudix ApnAase from M. tuberculosis and have subcloned it into a HisTag vector to optimize purification. This enzyme has been purified and characterized, and we are in the process of crystallization for structure determination. We have sent purified proteins to the Hauptmann Woodward Institute, and through screening, we have identified multiple crystallization conditions that we are optimizing. This research has been supported by an NIH AREA grant, a CUR-Goldwater Scholars Faculty Mentor Award, an ASBMB undergraduate research award, a RAS student research grant, and a RIT COS Emerson SURF.