Target-Based Screen Against a Periplasmic Serine Protease That Regulates Intrabacterial pH Homeostasis in Mycobacterium tuberculosis
| Autor: | Carl Nathan, Nan Zhao, Helene Botella, Jennifer L. Small, Christina Eberhart, Crystal M. Darby, Hugh Rosen, Benjamin F. Cravatt, Timothy P. Spicer, Xiuju Jiang, Peter Hodder, Sabine Ehrt, Daniel A. Bachovchin, Anna E Speers, Erin D. Anderson, Virneliz Fernandez-Vega, Kristin Burns-Huang, Dale L. Boger |
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| Rok vydání: | 2014 |
| Předmět: |
Serine Proteinase Inhibitors
medicine.medical_treatment Biochemistry Gene Expression Regulation Enzymologic Microbiology Mycobacterium tuberculosis Bacterial Proteins medicine Homeostasis Letters Phagosome chemistry.chemical_classification Serine protease Mycobacterium bovis Protease Molecular Structure biology Gene Expression Regulation Bacterial General Medicine Periplasmic space Hydrogen-Ion Concentration biology.organism_classification Benzoxazines 3. Good health Enzyme chemistry Molecular Probes Periplasm biology.protein Molecular Medicine Serine Proteases Genetic screen |
| Zdroj: | ACS Chemical Biology |
| ISSN: | 1554-8937 1554-8929 |
| DOI: | 10.1021/cb500746z |
| Popis: | Mycobacterium tuberculosis (Mtb) maintains its intrabacterial pH (pHIB) near neutrality in the acidic environment of phagosomes within activated macrophages. A previously reported genetic screen revealed that Mtb loses this ability when the mycobacterial acid resistance protease (marP) gene is disrupted. In the present study, a high throughput screen (HTS) of compounds against the protease domain of MarP identified benzoxazinones as inhibitors of MarP. A potent benzoxazinone, BO43 (6-chloro-2-(2′-methylphenyl)-4H-1,3-benzoxazin-4-one), acylated MarP and lowered Mtb’s pHIB and survival during incubation at pH 4.5. BO43 had similar effects on MarP-deficient Mtb, suggesting the existence of additional target(s). Reaction of an alkynyl-benzoxazinone, BO43T, with Mycobacterium bovis variant bacille Calmette-Guérin (BCG) followed by click chemistry with azido-biotin identified both the MarP homologue and the high temperature requirement A1 (HtrA1) homologue, an essential protein. Thus, the chemical probe identified through a target-based screen not only reacted with its intended target in the intact cells but also implicated an additional enzyme that had eluded a genetic screen biased against essential genes. |
| Databáze: | OpenAIRE |
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