High Throughput Crystallography of TB Drug Targets

Abstract

Tuberculosis (TB) infects one-third of the world population. Despite 50 years of available drug treatments, TB continues to increase at a significant rate. The failure to control TB stems in part from the expense of delivering treatment to infected individuals and from complex treatment regimens. Incomplete treatment has fueled the emergence of multi-drug resistant (MDR) strains of Mycobacterium tuberculosis (Mtb). Reducing non-compliance by reducing the duration of chemotherapy will have a great impact on TB control. The development of new drugs that either kill persisting organisms, inhibit bacilli from entering the persistent phase, or convert the persistent bacilli into actively growing cells susceptible to our current drugs will have a positive effect. We are taking a multidisciplinary approach that will identify and characterize new drug targets that are essential for persistent Mtb. Targets are exposed to a battery of analyses including microarray experiments, bioinformatics, and genetic techniques to prioritize potential drug targets from Mtb for structural analysis. Our core structural genomics pipeline works with the individual laboratories to produce diffraction quality crystals of targeted proteins, and structural analysis will be completed by the individual laboratories. We also have capabilities for functional analysis and the virtual ligand screening to identify novel inhibitors for target validation. Our overarching goals are to increase the knowledge of Mtb pathogenesis using the TB research community to drive structural genomics, particularly related to persistence, develop a central repository for TB research reagents, and discover chemical inhibitors of drug targets for future development of lead compounds.

M. tuberculosis Pantothenate Kinase: Dual Substrate Specificity and Unusual Changes in Ligand Locations

“…4,5 We have been exploring the structures and interactions of PanK from Mycobacterium tuberculosis (MtPanK) as part of a program in this laboratory [6][7][8][9][10][11][12] and an international effort involving structural studies on mycobacterial proteins. [13][14][15][16][17] The CoA complex of MtPanK has a structure very similar to that of the corresponding EcPanK complex. 7 However, unlike in the case of EcPanK, the structure observed in the MtPanK-CoA complex is retained in the complexes involving ADP and AMPPCP, another non-hydrolyzable analogue of ATP as well.…”

The Structure of the Arginine Repressor from Mycobacterium tuberculosis Bound with its DNA Operator and Co-repressor, L-Arginine

“…Therefore, a pressing goal is to increase our knowledge of Mtb infectivity and pathogenesis for the future development of more potent drugs and eventually a cure. 1 We have targeted one of the essential pathways in Mtb metabolism, that of arginine biosynthesis. 2 The arginine operon in Mtb includes seven genes required for arginine biosynthesis; it is regulated by a transcriptional factor also in the arginine operon, the arginine repressor (ArgR) that is the gene product of the open reading frame Rv1657.…”

The Molecular Structure of Epoxide Hydrolase B from Mycobacterium tuberculosis and Its Complex with a Urea-Based Inhibitor

“…The Mtb structural genomics consortium † , formed in 2000, aims to determine the three-dimensional structures of Mtb proteins that not only would advance the understanding of Mtb pathogenesis but also will help in the development of small-molecule inhibitors against potential drug targets using a structure-based drug design approach. 2 In fact, several promising small-molecule anti-TB compounds have emerged through this approach. 3 Sequencing of the complete genome of Mtb strain H37Rv (4.4 Mbp) has revealed the genes for at least six potential epoxide hydrolase (EH) proteins (open reading frames Rv0134, Rv1124, Rv1938, Rv2214c, Rv3617, and Rv3670) belonging to the virulence, detoxification, and adaptation functional category that encompasses 2.4% of the entire Mtb genome.…”