High-throughput screen identifies small molecule inhibitors targeting acetyltransferase activity of Mycobacterium tuberculosis GlmU

Rani, C.; Mehra, Rukmankesh; Chib, Reena; Wazir, Priya; Nargotra, Amit; Khan, Inshad Ali

doi:10.1016/j.tube.2015.06.003

Cited by 21 publications

(18 citation statements)

References 35 publications

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“…Rani et al found four compounds which showed an inhibitory effect on GlmU acetyltransferase activity, with IC 50 values ranging from 9 to 70 μM. Compounds 6624116 and 5655606 exhibited activity against drug-susceptible as well as drug-resistant Mtb (Rani et al, 2015). However, the detailed information of the action mechanism of these compounds was not reported.…”

Section: Introductionmentioning

confidence: 99%

The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c

Chen

Han

Yan

et al. 2019

Front. Cell. Infect. Microbiol.

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Mycobacterium tuberculosis bifunctional enzyme GlmU is a novel target for anti-TB drugs and is involved in glycosyl donor UDP-N-acetylglucosamine biosynthesis. Here, we found that TPSA (2-[5-(2-{[4-(2-thienyl)-2-pyrimidinyl]sulfanyl}acetyl)-2-thienyl]acetic acid) was a novel inhibitor for GlmU acetyltransferase activity (IC 50 : 5.3 µM). The interaction sites of GlmU and TPSA by molecular docking were confirmed by site-directed mutagenesis. TPSA showed an inhibitory effect on Mtb H37Ra growth and intracellular H37Ra in macrophage cells (MIC: 66.5 µM). To investigate why TPSA at a higher concentration (66.5 µM) was able to inhibit H37Ra growth, proteome and transcriptome of H37Ra treated with TPSA were analyzed. The expression of two methyltransferases MRA_0565 (Rv0558) and MRA_0567 (Rv0560c) were markedly increased. TPSA was pre-incubated with purified Rv0558 and Rv0560c in the presence of S-adenosylmethionine (methyl donor) respectively, resulting in its decreased inhibitory effect of GlmU on acetyltransferase activity. The inhibition of TPSA on growth of H37Ra with overexpressed Rv0558 and Rv0560c was reduced. These implied that methyltransferases could modify TPSA. The methylation of TPSA catalyzed by Rv0560c was subsequently confirmed by LC-MS. Therefore, TPSA as a GlmU acetyltransferase activity inhibitor may offer a structural basis for new anti-tuberculosis drugs. TPSA needs to be modified further by some groups to prevent its methylation by methyltransferases.

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Section: Introductionmentioning

confidence: 99%

The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c

Chen

Han

Yan

et al. 2019

Front. Cell. Infect. Microbiol.

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“…The limited number of new anti-mycobacterial agents approved for therapy and the wide variety of Mtb intrinsic and acquired drug resistance mechanisms to the available drugs have contributed to an increased effort to repurpose the use of antibiotics that are not commonly used in anti-TB therapy and to find suitable synergistic antibiotic combinations for effective treatment of life-risk TB (Mainardi et al., 2011; Wong et al., 2013; Keener, 2014; Diacon et al., 2016). Recent studies have uncovered the possibility of targeting the mycobacterial PG biosynthesis and degradation as an alternative option for anti-TB therapy (Tomasic et al., 2010; Trunkfield et al., 2010; Li et al., 2011; Rana et al., 2014; Ling et al., 2015; Rani et al., 2015; Rullas et al., 2015; Tran et al., 2017). In addition, several observations suggest that inhibition of PG synthesis by transpeptidase inhibitors such as the carbapenems or glycopeptide antibiotics could synergize with other CW inhibitors and increase their efficacy (Figures 1 and 2; Hugonnet et al., 2009; Kumar et al., 2012; Kieser et al., 2015; Schubert et al., 2017).…”

Section: Discussionmentioning

confidence: 99%

“…The functional resemblance of the GlmU uridyltransferase with human enzymes (Peneff et al., 2001) turns this enzyme into an unsuitable target (Rani and Khan, 2016). However, the lack of Glc N -1-P from mammals makes the acetyltransferase domain a promising target, and different substrate analogs of Glc N -1-P have been designed and shown to exhibit an inhibitory effect against GlmU by blocking synthesis of UDP-Glc N Ac (Figure 1; Li et al., 2011; Tran et al., 2013; Rani et al., 2015). The sequential MurA-F ligase pathway involves the formation of the UDP- N -acetylmuramic acid (UDP-Mur N Ac)-pentapeptide.…”

Section: Mycobacterial Pg Assembly Enzymes As Targets For Antibioticsmentioning

confidence: 99%

Revisiting Anti-tuberculosis Therapeutic Strategies That Target the Peptidoglycan Structure and Synthesis

2019

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Tuberculosis (TB), which is caused by Mycobacterium tuberculosis (Mtb), is one of the leading cause of death by an infectious diseases. The biosynthesis of the mycobacterial cell wall (CW) is an area of increasing research significance, as numerous antibiotics used to treat TB target biosynthesis pathways of essential CW components. The main feature of the mycobacterial cell envelope is an intricate structure, the mycolyl-arabinogalactan-peptidoglycan (mAGP) complex responsible for its innate resistance to many commonly used antibiotics and involved in virulence. A hallmark of mAGP is its unusual peptidoglycan (PG) layer, which has subtleties that play a key role in virulence by enabling pathogenic species to survive inside the host and resist antibiotic pressure. This dynamic and essential structure is not a target of currently used therapeutics as Mtb is considered naturally resistant to most β-lactam antibiotics due to a highly active β-lactamase (BlaC) that efficiently hydrolyses many β-lactam drugs to render them ineffective. The emergence of multidrug- and extensive drug-resistant strains to the available antibiotics has become a serious health threat, places an immense burden on health care systems, and poses particular therapeutic challenges. Therefore, it is crucial to explore additional Mtb vulnerabilities that can be used to combat TB. Remodeling PG enzymes that catalyze biosynthesis and recycling of the PG are essential to the viability of Mtb and are therefore attractive targets for novel antibiotics research. This article reviews PG as an alternative antibiotic target for TB treatment, how Mtb has developed resistance to currently available antibiotics directed to PG biosynthesis, and the potential of targeting this essential structure to tackle TB by attacking alternative enzymatic activities involved in Mtb PG modifications and metabolism.

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“…Target-based small-molecule screenings are a valuable and widely used approach for lead generation in drug discovery. In traditional screenings compounds are tested against purified enzymes, with the goal of detecting enzyme inhibition 29 , 30 , activation 31 , or binding to the target protein 32 , while other screenings test for secondary ligands that only bind after the first ligand binds 33 , 34 . Despite their undisputed usefulness, traditional screens fail to detect compounds displaying less common or peculiar effects, such as influencing the efficacy or affinity of a known effector, or altering effector–enzyme cooperativity.…”

Section: Discussionmentioning

confidence: 99%

Uncoupling conformational states from activity in an allosteric enzyme

et al. 2017

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ATP-phosphoribosyltransferase (ATP-PRT) is a hexameric enzyme in conformational equilibrium between an open and seemingly active state and a closed and presumably inhibited form. The structure-function relationship of allosteric regulation in this system is still not fully understood. Here, we develop a screening strategy for modulators of ATP-PRT and identify 3-(2-thienyl)-l-alanine (TIH) as an allosteric activator of this enzyme. Kinetic analysis reveals co-occupancy of the allosteric sites by TIH and l-histidine. Crystallographic and native ion-mobility mass spectrometry data show that the TIH-bound activated form of the enzyme closely resembles the inhibited l-histidine-bound closed conformation, revealing the uncoupling between ATP-PRT open and closed conformations and its functional state. These findings suggest that dynamic processes are responsible for ATP-PRT allosteric regulation and that similar mechanisms might also be found in other enzymes bearing a ferredoxin-like allosteric domain.

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High-throughput screen identifies small molecule inhibitors targeting acetyltransferase activity of Mycobacterium tuberculosis GlmU

Cited by 21 publications

References 35 publications

The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c

The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c

Revisiting Anti-tuberculosis Therapeutic Strategies That Target the Peptidoglycan Structure and Synthesis

Uncoupling conformational states from activity in an allosteric enzyme

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