Differential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platform

Theron, AnjoRoth, Robyn LHoppe, HParkinson, CVan der Westhuyzen, Christiaan WStoychev, Stoyan HWiid, IPietersen, RDBaker, BKenyon, CP2017-11-202017-11-202017-10Theron, A. et al. 2017. Differential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platform. PLOS ONE, vol 12(10): e0185068. https://doi.org/10.1371/journal.pone.01850681932-6203http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0185068doi.org/10.1371/journal.pone.0185068http://hdl.handle.net/10204/9802Copyright: 2017 The Authors. This is an Open Access article.Glutamine synthetase is a ubiquitous central enzyme in nitrogen metabolism that is controlled by up to four regulatory mechanisms, including adenylylation of some or all of the twelve subunits by adenylyl transferase. It is considered a potential therapeutic target for the treatment of tuberculosis, being essential for the growth of Mycobacterium tuberculosis, and is found extracellularly only in the pathogenic Mycobacterium strains. Human glutamine synthetase is not regulated by the adenylylation mechanism, so the adenylylated form of bacterial glutamine synthetase is of particular interest. Previously published reports show that, when M. tuberculosis glutamine synthetase is expressed in Escherichia coli, the E. coli adenylyl transferase does not optimally adenylylate the M. tuberculosis glutamine synthetase. Here, we demonstrate the production of soluble adenylylated M. tuberulosis glutamine synthetase in E. coli by the co-expression of M. tuberculosis glutamine synthetase and M. tuberculosis adenylyl transferase. The differential inhibition of adenylylated M. tuberulosis glutamine synthetase and deadenylylated M. tuberulosis glutamine synthetase by ATP based scaffold inhibitors are reported. Compounds selected on the basis of their enzyme inhibition were also shown to inhibit M. tuberculosis in the BACTEC 460TB™ assay as well as the intracellular inhibition of M. tuberculosis in a mouse bone-marrow derived macrophage assay.enGlutamine synthetaseMycobacterium tuberculosisDifferential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platformArticleTheron, A., Roth, R. L., Hoppe, H., Parkinson, C., Van der Westhuyzen, C. W., Stoychev, S. H., ... Kenyon, C. (2017). Differential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platform. http://hdl.handle.net/10204/9802Theron, Anjo, Robyn L Roth, H Hoppe, C Parkinson, Christiaan W Van der Westhuyzen, Stoyan H Stoychev, I Wiid, RD Pietersen, B Baker, and CP Kenyon "Differential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platform." (2017) http://hdl.handle.net/10204/9802Theron A, Roth RL, Hoppe H, Parkinson C, Van der Westhuyzen CW, Stoychev SH, et al. Differential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platform. 2017; http://hdl.handle.net/10204/9802.TY - Article AU - Theron, Anjo AU - Roth, Robyn L AU - Hoppe, H AU - Parkinson, C AU - Van der Westhuyzen, Christiaan W AU - Stoychev, Stoyan H AU - Wiid, I AU - Pietersen, RD AU - Baker, B AU - Kenyon, CP AB - Glutamine synthetase is a ubiquitous central enzyme in nitrogen metabolism that is controlled by up to four regulatory mechanisms, including adenylylation of some or all of the twelve subunits by adenylyl transferase. It is considered a potential therapeutic target for the treatment of tuberculosis, being essential for the growth of Mycobacterium tuberculosis, and is found extracellularly only in the pathogenic Mycobacterium strains. Human glutamine synthetase is not regulated by the adenylylation mechanism, so the adenylylated form of bacterial glutamine synthetase is of particular interest. Previously published reports show that, when M. tuberculosis glutamine synthetase is expressed in Escherichia coli, the E. coli adenylyl transferase does not optimally adenylylate the M. tuberculosis glutamine synthetase. Here, we demonstrate the production of soluble adenylylated M. tuberulosis glutamine synthetase in E. coli by the co-expression of M. tuberculosis glutamine synthetase and M. tuberculosis adenylyl transferase. The differential inhibition of adenylylated M. tuberulosis glutamine synthetase and deadenylylated M. tuberulosis glutamine synthetase by ATP based scaffold inhibitors are reported. Compounds selected on the basis of their enzyme inhibition were also shown to inhibit M. tuberculosis in the BACTEC 460TB™ assay as well as the intracellular inhibition of M. tuberculosis in a mouse bone-marrow derived macrophage assay. DA - 2017-10 DB - ResearchSpace DP - CSIR KW - Glutamine synthetase KW - Mycobacterium tuberculosis LK - https://researchspace.csir.co.za PY - 2017 SM - 1932-6203 T1 - Differential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platform TI - Differential inhibition of adenylylated and deadenylylated forms of M. tuberculosis glutamine synthetase as a drug discovery platform UR - http://hdl.handle.net/10204/9802 ER -