Grant: $50,054 - Department of Health and Human Services - Aug. 14, 2009
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Award Description: Mycobacterium tuberculosis (Mtb) is the leading cause of pathogenic infectious disease mortality worldwide. Roughly one-third of the world?s population is infected with Mtb and nearly two million people die from this disease per year. Thus, there is a critical need for the development of new therapeutic agents for the treatment of Mtb. These new therapies will ideally target new pathways and will posses mechanisms of action that are divergent from current Mtb drugs. Adenylating enzymes of Mtb have recently emerged as attractive targets for therapeutic drug development. Mtb contains more than 100 adenylating enzymes (AEs), which play vital roles in many aspects of the metabolism of the pathogen. AEs have been implicated in lipid metabolism, biosynthesis of siderophores and cofactors, as well as protein and DNA synthesis. Two families of Mtb AEs have attracted the attention of the Aldrich lab for their direct role in the virulence of Mtb: aryl acid adenylating enzymes (AAAEs) and fatty acid adenylating enzymes (FadDs). These two enzyme families share a common protein fold that bind the acyl-adenylate intermediate at the interface of the N¬- and C- terminal domains. Inhibitors that individually target each of these AEs possess impressive anti-tubercular activity. Activity based protein profiling (ABPP) is a powerful proteomic tool that uses chemical probes to study large enzyme families. ABPP has been instrumental in identifying novel inhibitors of whole enzyme classes as well as uncovering unintended off-targets of enzyme inhibitors. Surprisingly, no ABPP chemical probes have been described for the adenylating enzyme family. Since AEs have been shown to be important in Mtb growth and virulence, ABPP chemical probes are needed to study this significant enzyme class. Therefore, the objectives of this application are to develop chemical probes to study Mtb AEs and their inhibition. We plan to accomplish this objective by pursuing the following two specific aims: Specific Aim 1: Develop Activity Based Protein Profiling Probes for Mtb aryl acid adenylating enzymes. The Aldrich lab has developed a potent inhibitor of an Mtb aryl acid adenylating enzyme (AAAE), MbtA. The studies in this aim are directed at developing an activity based chemical probe to profile the inhibition of this particular AAAE. Ultimately, our ABPP probe will aid in identifying off-targets of this inhibitor. Specific Aim 2: Develop an Activity Based Protein Profiling Platform for FadD Inhibitor Screening. Work in this aim will involve the identification of protein targets of a FadD inhibitor. To accomplish this, an activity based chemical probe will be designed to target and covalently label multiple FadDs. Inhibitors will then be screened for their ability to displace the active site probe using gel electrophoresis and LCMS-MS analysis. This study will provide insight into inhibitor potency and selectivity towards multiple FadDs. These studies are innovative in that they will yield chemical tools to study Mtb adenylating enzymes. Additionally, these probes will ultimately help to illuminate the molecular target of two potent antitubercular agents. It is expected that upon completion of specific aim 1, we will have a thorough knowledge of the protein off-targets of a Mtb inhibitor. Additionally, the developed probe will be available for studying the inhibition of AAAEs other than MbtA. Upon completion of specific aim 2, we will have identified the protein target of a FadD inhibitor. Additionally, a platform will be in place that can be rapidly used to screen FadD inhibitors for their ability to disrupt and inhibit multiple FadDs. Taken together, these results are expected to validate the use of inhibitors that target novel Mtb biosynthetic pathways as treatments for tuberculosis.
Project Description: As defined in the award description field.
Jobs Summary: No jobs reported. (Total jobs reported: 0)
Project Status: Less Than 50% Completed
This award's data was last updated on Aug. 14, 2009. Help expand these official descriptions using the wiki below.