Adenosine Tetraphosphoadenosine Drives a Continuous ATP-Release Assay for Aminoacyl-tRNA Synthetases and Other Adenylate-Forming Enzymes

Lloyd, Adrian J.; Potter, Nicola J; Fishwick, Colin W. G.; Roper, David I.; Dowson, Christopher G.

doi:10.1021/cb400248f

Cited by 10 publications

(14 citation statements)

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“…In addition, in some situations, several different amino acids are able to bind to non-cognate aaRSs, requiring an in vivo editing function allowing for the possibility of exploiting this feature for future antimicrobial discovery 6 . For example, the amino acid serine is able to bind alanyl-tRNA synthetase (AlaRS) and threonyl-tRNA synthetase (ThrRS) in addition to its cognate seryl-tRNA synthetase (SerRS) 7 . This incorrect binding is rectified in nature by numerous proofreading mechanisms 6,8 .…”

Section: Introductionmentioning

confidence: 99%

“…Mupirocin targets the IleRS enzyme and utilises a hydrophobic "tail" in addition to an aminoacyl adenylate warhead to bind to its target 11 . By contrast to many antibiotics in clinical use, seryl sulfamoyl adenosine (SerSA, 1) can bind and inhibit AlaRS and ThrRS in addition to SerRS and hence is a multi-targeting inhibitor 7,12 . It can be predicted therefore that SerSA would require mutations in several of these enzymes before a resistance phenotype could be conferred.…”

Section: Introductionmentioning

confidence: 99%

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Structure-guided enhancement of selectivity of chemical probe inhibitors targeting bacterial seryl-tRNA synthetase

Cain¹,

Salimaraj²,

Punekar³

et al. 2019

Preprint

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Aminoacyl-tRNA synthetases are ubiquitous and essential enzymes for protein synthesis and also a variety of other metabolic processes, especially in bacterial species. Bacterial aminoacyl-tRNA synthetases represent attractive and validated targets for antimicrobial drug discovery if issues of prokaryotic versus eukaryotic selectivity and antibiotic resistance generation can be addressed. We have determined high resolution X-ray crystal structures of the Escherichia coli and Staphylococcus aureus seryl-tRNA synthetases in complex with aminoacyl adenylate analogues and applied a structure-based drug discovery approach to explore and identify a series of small molecule inhibitors that selectively inhibit bacterial seryl-tRNA synthetases with greater than two orders of magnitude compared to their human homologue, demonstrating a route to selective chemical inhibition of these bacterial targets.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Structure-guided enhancement of selectivity of chemical probe inhibitors targeting bacterial seryl-tRNA synthetase

Cain¹,

Salimaraj²,

Punekar³

et al. 2019

Preprint

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“…butyl N-[(2S)-1-[(([(3aR, 4R, 6R, 6aR)-6-(6-amino-2-chloro-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-2H-furo[3,4-d][1,3]dioxol-4yl]methoxy)sulfonyl)amino]-3-(benzyloxy)-1oxopropan-2-yl]carbamate (0.30 g, 0.43 mmol) to afford the desired product as a colourless powder(50.4 mg, 0.11 mmol, 25%) m.p. : 121.3 °C (Decomp); Rf: Baseline (9:1 DCM-MeOH); δH (500 MHz, DMSO-d6): 8.04 (1H, s, 6-HAr), 7.65 (2H, brs, NH2), 5.97 (1H, d, J 8.2, 2-HFuryl), 4.60 (1H, apps, 3-HFuryl), 4.31 (1H, dd, J 10.1 and 5.9, CH2*O), 4.19-4.15 (2H, m, CH2*O and 4-HFuryl), 4.13 (1H, app s, 5-HFuryl), 3.81 (1H, dd, J 10.2 and 7.3, CH2*chiral), 3.76 (1H, d, J 7.3, CHChiral), 3.65 (1H, dd, J 10.2 and 7.3, CH2*chiral); δC (125 MHz, DMSO-d6): 172.1 (C=O), 157.8 (C4Ar), 154.1 (C2Ar), 151.0 (C9Ar), 140.1 (C6Ar), 119.1 (C10Ar), 89.4 (C2Furyl), 84.4 (C5Furyl), 73.9 (C3Furyl), 71.5 (C4Furyl), 70.3 (CH2O), 64.4 (CH2Chiral), 56.9 (CHChiral); νmax/ cm -1 (solid): 3321, 3125, 2784, 1673, 1592, 1358; m/z (ES): No mass ion found [α]D = 28.8° (c 0.1, MeOH).Preparation of 2'3'-O-Isopropylidene-2-iodoadenosine2-iodoadenosine (1.00 g, 2.54 mmol, 1.0 equiv.)…”

mentioning

confidence: 99%

“…-furo[3,4-d][1,3]dioxol-4yl]methyl sulfamate (0.20 g, 0.43 mmol) to afford the desired product as a colourless glassy solid (0.21 g, 0.29 mmol, 67%) m.p.m, Chiral CH2), 1.59 (3H, s, CH3a), 1.39 (9H, s, CH3 × 3), 1.35 (3H, s, CH3b); δC (125 MHz, DMSO-d6): 173.5 (C=ONH), 158.7 (C2Ar), 156.8 (C4Ar), 156.4 (C=O BOC), 150.6 (C8Ar), 140.0 (C6Ar), 138.5 (C1'Ar and C1Bzl), 130.4 (C4'Ar), 129. C3Bzl, C4Bzl, C5Bzl, C6Bzl, …”

mentioning

confidence: 99%

Structure-Guided Enhancement of Selectivity of Chemical Probe Inhibitors Targeting Bacterial Seryl-tRNA Synthetase

et al. 2019

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Aminoacyl-tRNA synthetases are ubiquitous and essential enzymes for protein synthesis and also a variety of other metabolic processes, especially in bacterial species. Bacterial aminoacyl-tRNA synthetases represent attractive and validated targets for antimicrobial drug discovery if issues of prokaryotic versus eukaryotic selectivity and antibiotic resistance generation can be addressed. We have determined high-resolution X-ray crystal structures of the Escherichia coli and Staphylococcus aureus seryl-tRNA synthetases in complex with aminoacyl adenylate analogues and applied a structure-based drug discovery approach to explore and identify a series of small molecule inhibitors that selectively inhibit bacterial seryl-tRNA synthetases with greater than 2 orders of magnitude compared to their human homologue, demonstrating a route to the selective chemical inhibition of these bacterial targets.

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“…It has been proposed that the resistance liabilities associated with aaRS inhibitors could be overcome with an inhibitor capable of targeting two or more aaRS enzymes simultaneously (1,2,6); an equivalent effect could be achieved with a cocktail of two or more aaRS inhibitors delivered in combination. This proposal is supported by the multitarget hypothesis, which states that antibacterial agents for which resistance is not readily selected by mutation usually act on more than one cellular target (7).…”

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confidence: 99%

Targeting Multiple Aminoacyl-tRNA Synthetases Overcomes the Resistance Liabilities Associated with Antibacterial Inhibitors Acting on a Single Such Enzyme

Randall

Rasiņa

Jirgensons

et al. 2016

Antimicrob Agents Chemother

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bBacterial aminoacyl-tRNA synthetases (aaRSs) represent promising antibacterial drug targets. Unfortunately, the aaRS inhibitors that have to date reached clinical trials are subject to rapid resistance development through mutation, a phenomenon that limits their potential clinical utility. Here, we confirm the intuitively correct idea that simultaneous targeting of two different aaRS enzymes prevents the emergence of spontaneous bacterial resistance at high frequency, a finding that supports the development of multitargeted anti-aaRS therapies.T he aminoacyl-tRNA synthetase (aaRS) enzymes possess several features that render them promising prospects as broadspectrum antibacterial drug targets; they are essential for viability, are found in all bacterial pathogens, and are in many cases sufficiently structurally distinct from their eukaryotic counterparts to allow selective targeting (1, 2). Furthermore, chemical and clinical validation exists for these enzymes as useful targets for antibacterial chemotherapy (1). However, despite the potential promise of this family of targets, only a single aaRS inhibitor with a relatively limited indication has been approved for the management of bacterial infection. Mupirocin (MUP), an inhibitor of isoleucyltRNA synthetase, is a topical agent deployed for nasal decolonization of Staphylococcus aureus and for the treatment of superficial skin infection (3).Unfortunately, as with other antibacterial agents that act on a single enzyme target, aaRS inhibitors possess an intrinsic resistance liability (4). Mutants resistant to aaRS inhibitors are selected at a high frequency in bacterial populations (ϳ10 Ϫ7 ), typically as a result of point mutations within the gene encoding the drug target that lead to alteration of the latter in a manner that negatively impacts inhibitor binding (1). This liability, while manageable in the context of aaRS inhibitors such as MUP that are applied topically at concentrations sufficiently high to prevent or mitigate resistance, presents a problem for the development of aaRS inhibitors for systemic treatment of more serious bacterial disease. Indeed, GlaxoSmithKline halted phase II clinical trials of the leucyltRNA synthetase inhibitor GSK2251052 for the treatment of complicated urinary tract infections in adults following the emergence of mutants of Escherichia coli that were resistant to the drug in 3 of 14 patients within 2 days of administration (5).It has been proposed that the resistance liabilities associated with aaRS inhibitors could be overcome with an inhibitor capable of targeting two or more aaRS enzymes simultaneously (1, 2, 6); an equivalent effect could be achieved with a cocktail of two or more aaRS inhibitors delivered in combination. This proposal is supported by the multitarget hypothesis, which states that antibacterial agents for which resistance is not readily selected by mutation usually act on more than one cellular target (7). By targeting two or more aaRS enzymes simultaneously, a situation is created in which the likeliho...

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Adenosine Tetraphosphoadenosine Drives a Continuous ATP-Release Assay for Aminoacyl-tRNA Synthetases and Other Adenylate-Forming Enzymes

Cited by 10 publications

References 32 publications

Structure-guided enhancement of selectivity of chemical probe inhibitors targeting bacterial seryl-tRNA synthetase

Structure-guided enhancement of selectivity of chemical probe inhibitors targeting bacterial seryl-tRNA synthetase

Structure-Guided Enhancement of Selectivity of Chemical Probe Inhibitors Targeting Bacterial Seryl-tRNA Synthetase

Targeting Multiple Aminoacyl-tRNA Synthetases Overcomes the Resistance Liabilities Associated with Antibacterial Inhibitors Acting on a Single Such Enzyme

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