title: Target-based identification of whole-cell active inhibitors of biotin biosynthesis in Mycobacterium tuberculosis.
abstract: Biotin biosynthesis is essential for survival and persistence of Mycobacterium tuberculosis (Mtb) in vivo. The aminotransferase BioA, which catalyzes the antepenultimate step in the biotin pathway, has been established as a promising target due to its vulnerability to chemical inhibition. We performed high-throughput screening (HTS) employing a fluorescence displacement assay and identified a diverse set of potent inhibitors including many diversity-oriented synthesis (DOS) scaffolds. To efficiently select only hits targeting biotin biosynthesis, we then deployed a whole-cell counterscreen in biotin-free and biotin-containing medium against wild-type Mtb and in parallel with isogenic bioA Mtb strains that possess differential levels of BioA expression. This counterscreen proved crucial to filter out compounds whose whole-cell activity was off target as well as identify hits with weak, but measurable whole-cell activity in BioA-depleted strains. Several of the most promising hits were cocrystallized with BioA to provide a framework for future structure-based drug design efforts.
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Sae Woong
authors.initials: SW
authors.lastname: Park
authors.affiliation: The Broad Institute Probe Development Center, Cambridge, MA 02142, USA.
authors.forename: Dominick E
authors.initials: DE
authors.lastname: Casalena
authors.affiliation: Center for Drug Design, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Wilson
authors.affiliation: Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Ran
authors.initials: R
authors.lastname: Dai
authors.affiliation: The Broad Institute Probe Development Center, Cambridge, MA 02142, USA.
authors.forename: Partha P
authors.initials: PP
authors.lastname: Nag
authors.affiliation: Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.affiliation: Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-6604, USA.
authors.forename: Jim P
authors.initials: JP
authors.lastname: Boyce
authors.affiliation: The Broad Institute Probe Development Center, Cambridge, MA 02142, USA.
authors.forename: Joshua A
authors.initials: JA
authors.lastname: Bittker
authors.affiliation: The Broad Institute Probe Development Center, Cambridge, MA 02142, USA.
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Schreiber
authors.affiliation: Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Barry C
authors.initials: BC
authors.lastname: Finzel
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA. Electronic address: dis2003@med.cornell.edu.
authors.forename: Dirk
authors.initials: D
authors.lastname: Schnappinger
authors.affiliation: Center for Drug Design, University of Minnesota, Minneapolis, MN 55455, USA; Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: aldri015@umn.edu.
authors.forename: Courtney C
authors.initials: CC
authors.lastname: Aldrich
mesh_terms: D000995:Antitubercular Agents / Q000737:chemistry / Q000494:pharmacology; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D001665:Binding Sites; D001710:Biotin / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D002151:Calorimetry; D018360:Crystallography, X-Ray; D015195:Drug Design; D057166:High-Throughput Screening Assays; D006860:Hydrogen Bonding; D008826:Microbial Sensitivity Tests; D056004:Molecular Dynamics Simulation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D000637:Transaminases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D001710:Biotin; D000637:Transaminases; C102100:BioA protein, bacteria