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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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2014
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.
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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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