All ETDs from UAB

Advisory Committee Chair

Adrie Jc Steyn

Advisory Committee Members

William Benjamin

Michael Niederweis

Robert Reynolds

Janet Yother

Document Type

Dissertation

Date of Award

2010

Degree Name by School

Doctor of Philosophy (PhD) Heersink School of Medicine

Abstract

Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis (TB) and infects nearly one-third of the world's population. The same TB chemotherapeutic regimen has been administered for more than 50 years, and can be improved by developing innovative methods to discover new anti-TB drugs. This work describes a newly established high-throughput screen (HTS) platform technology based on Mycobacterial-Protein Fragment Complementation (M-PFC) to identify small molecule inhibitors of protein-protein interactions in mycobacteria. M-PFC HTS employs both whole cell and target-based approaches, and offers flexibility in selecting an appropriate protein interaction target for HTS. Selectable drug targets can include interactions of essential regulators (IdeR dimerization), enzymatic complexes (LeuCD), secretory antigens (Cfp10-Esat6), and signaling pathways (DevR dimerization). A proof-of-concept quantitative HTS (qHTS) of 3,600 small molecule compounds was performed on DevR-DevR interaction, a target chosen because of its potential involvement in Mtb persistence and the need for effective drugs against latent TB. The calculated Z'-factor was consistently ≥ 0.8, indicating a robust and reproducible assay. Completion of the proof-of-concept screen allowed for the identification of advantages and disadvantages in the current assay design, where improvements made will further pioneer M-PFC-based applications in a large-scale HTS format.

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