title: A screen to identify small molecule inhibitors of protein-protein interactions in mycobacteria.
abstract: Despite extensive efforts in tuberculosis (TB) drug research, very few novel inhibitors have been discovered. This issue emphasizes the need for innovative methods to discover new anti-TB drugs. In this study, we established a new high-throughput screen (HTS) platform technology that differs from traditional TB drug screens because it utilizes Mycobacterial-Protein Fragment Complementation (M-PFC) to identify small molecule inhibitors of protein-protein interactions in mycobacteria. Several examples of protein-protein interactions were tested with M-PFC to highlight the diversity of selectable drug targets that could be used for screening. These included interactions of essential regulators (IdeR dimerization), enzymatic complexes (LeuCD), secretory antigens (Cfp10-Esat6), and signaling pathways (DevR dimerization). The feasibility of M-PFC in a HTS platform setting was tested by performing a proof-of-concept quantitative HTS of 3,600 small molecule compounds on DevR-DevR interaction, which was chosen because of its strong implications in Mycobacterium tuberculosis 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.
authors.affiliation: The Department of Microbiology, University of Alabama at Birmingham, USA.
authors.forename: Deborah
authors.initials: D
authors.lastname: Mai
authors.forename: Jennifer
authors.initials: J
authors.lastname: Jones
authors.forename: John W
authors.initials: JW
authors.lastname: Rodgers
authors.forename: John L
authors.initials: JL
authors.lastname: Hartman
authors.forename: Olaf
authors.initials: O
authors.lastname: Kutsch
authors.forename: Adrie J C
authors.initials: AJ
authors.lastname: Steyn
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D025941:Protein Interaction Mapping / Q000379:methods*
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins