Abstract
The diadenosine polyphosphatases (ApnAases) / mRNA decapping enzymes are a family of enzymes within the Nudix hydrolase superfamily. 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. An ApnAases / mRNA decapping enzyme from Mycobacterium tuberculosis has been characterized in our laboratory and a homolog to this enzyme has been uncovered in Mycobacterium leprae. We have cloned and expressed the M. leprae homolog and have determined that it is an ApnAase. While the enzyme has good expression, it is rather insoluble. We have lowered the expression temperature, lowered the concentration of IPTG, incorporated GroESL, and used a Rosetta cell line to try to increase solubility, but without much effect. We are still working to increase solubility, before we purify and further characterize the enzyme. If these Mycobacteria ApnAases / mRNA decapping enzymes are found to be involved in invasiveness and thus in virulence, then these enzymes could be potential novel antibiotic targets in M. tuberculosis and M. leprae. This research was supported by a NIH AREA Grant and a RIT Graduate Student Grant.
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