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Target-Based Screen Against a Periplasmic Serine Protease That Regulates Intrabacterial pH Homeostasis in Mycobacterium tuberculosis
[Image: see text] Mycobacterium tuberculosis (Mtb) maintains its intrabacterial pH (pH(IB)) 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...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American
Chemical Society
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4340348/ https://www.ncbi.nlm.nih.gov/pubmed/25457457 http://dx.doi.org/10.1021/cb500746z |
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author | Zhao, Nan Darby, Crystal M. Small, Jennifer Bachovchin, Daniel A. Jiang, Xiuju Burns-Huang, Kristin E. Botella, Helene Ehrt, Sabine Boger, Dale L. Anderson, Erin D. Cravatt, Benjamin F. Speers, Anna E. Fernandez-Vega, Virneliz Hodder, Peter S. Eberhart, Christina Rosen, Hugh Spicer, Timothy P. Nathan, Carl F. |
author_facet | Zhao, Nan Darby, Crystal M. Small, Jennifer Bachovchin, Daniel A. Jiang, Xiuju Burns-Huang, Kristin E. Botella, Helene Ehrt, Sabine Boger, Dale L. Anderson, Erin D. Cravatt, Benjamin F. Speers, Anna E. Fernandez-Vega, Virneliz Hodder, Peter S. Eberhart, Christina Rosen, Hugh Spicer, Timothy P. Nathan, Carl F. |
author_sort | Zhao, Nan |
collection | PubMed |
description | [Image: see text] Mycobacterium tuberculosis (Mtb) maintains its intrabacterial pH (pH(IB)) 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 pH(IB) 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. |
format | Online Article Text |
id | pubmed-4340348 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American
Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-43403482015-12-02 Target-Based Screen Against a Periplasmic Serine Protease That Regulates Intrabacterial pH Homeostasis in Mycobacterium tuberculosis Zhao, Nan Darby, Crystal M. Small, Jennifer Bachovchin, Daniel A. Jiang, Xiuju Burns-Huang, Kristin E. Botella, Helene Ehrt, Sabine Boger, Dale L. Anderson, Erin D. Cravatt, Benjamin F. Speers, Anna E. Fernandez-Vega, Virneliz Hodder, Peter S. Eberhart, Christina Rosen, Hugh Spicer, Timothy P. Nathan, Carl F. ACS Chem Biol [Image: see text] Mycobacterium tuberculosis (Mtb) maintains its intrabacterial pH (pH(IB)) 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 pH(IB) 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. American Chemical Society 2014-12-02 2015-02-20 /pmc/articles/PMC4340348/ /pubmed/25457457 http://dx.doi.org/10.1021/cb500746z Text en Copyright © 2014 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes. |
spellingShingle | Zhao, Nan Darby, Crystal M. Small, Jennifer Bachovchin, Daniel A. Jiang, Xiuju Burns-Huang, Kristin E. Botella, Helene Ehrt, Sabine Boger, Dale L. Anderson, Erin D. Cravatt, Benjamin F. Speers, Anna E. Fernandez-Vega, Virneliz Hodder, Peter S. Eberhart, Christina Rosen, Hugh Spicer, Timothy P. Nathan, Carl F. Target-Based Screen Against a Periplasmic Serine Protease That Regulates Intrabacterial pH Homeostasis in Mycobacterium tuberculosis |
title | Target-Based Screen Against a Periplasmic Serine Protease
That Regulates Intrabacterial pH Homeostasis in Mycobacterium
tuberculosis |
title_full | Target-Based Screen Against a Periplasmic Serine Protease
That Regulates Intrabacterial pH Homeostasis in Mycobacterium
tuberculosis |
title_fullStr | Target-Based Screen Against a Periplasmic Serine Protease
That Regulates Intrabacterial pH Homeostasis in Mycobacterium
tuberculosis |
title_full_unstemmed | Target-Based Screen Against a Periplasmic Serine Protease
That Regulates Intrabacterial pH Homeostasis in Mycobacterium
tuberculosis |
title_short | Target-Based Screen Against a Periplasmic Serine Protease
That Regulates Intrabacterial pH Homeostasis in Mycobacterium
tuberculosis |
title_sort | target-based screen against a periplasmic serine protease
that regulates intrabacterial ph homeostasis in mycobacterium
tuberculosis |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4340348/ https://www.ncbi.nlm.nih.gov/pubmed/25457457 http://dx.doi.org/10.1021/cb500746z |
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