Preliminary X-ray crystallographic analysis of ornithine acetyltransferase (Rv1653) from<i>Mycobacterium tuberculosis</i>

Sankaranarayanan, R.; Garen, Craig R.; Cherney, M.M.; Yuan, Marshall; Lee, C.; James, Michael N.G.

doi:10.1107/s1744309109000360

Cited by 4 publications

(1 citation statement)

References 22 publications

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“…29 The native orthorhombic crystals were grown from a solution containing a protein concentration of 13.5 mg/ml in 2.5 mM Hepes buffer (pH 7.5) and a precipitant solution consisting of 0.02 M MgCl 2 and 22% polyacrylic acid 5100 dissolved in 0.1 M Hepes (pH 7.5). The native crystals in complex with ORN were prepared from an overnight soaking of native crystals in a solution of 3 mM ORN in mother liquor.…”

Section: Crystallizationmentioning

confidence: 99%

The Molecular Structure of Ornithine Acetyltransferase from Mycobacterium tuberculosis Bound to Ornithine, a Competitive Inhibitor

Sankaranarayanan

Cherney

Garen

et al. 2010

Journal of Molecular Biology

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Section: Crystallizationmentioning

confidence: 99%

The Molecular Structure of Ornithine Acetyltransferase from Mycobacterium tuberculosis Bound to Ornithine, a Competitive Inhibitor

Sankaranarayanan

Cherney

Garen

et al. 2010

Journal of Molecular Biology

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Analysis of metabolic pathways in mycobacteria to aid drug-target identification

Bannerman

Vedithi

Júlvez

et al. 2019

Preprint

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Full Title: Comparative analyses of metabolic pathways in mycobacteria: Insights into the identification and characterisation of new drug targets in M. tuberculosis, M. leprae and M. abscessus Abstract Three related mycobacteria are the cause of widespread infections in man and are the focus of intense research and drug-discovery efforts in the face of growing antimicrobial resistance.Mycobacterium tuberculosis, the causative agent of tuberculosis, is currently one of the top ten causes of death in the world according to WHO; M. abscessus, a group of non-tuberculous mycobacteria causes lung infections and other opportunistic infections in humans; and M. leprae, the causative agent of leprosy, remains endemic in tropical countries. There is an urgent need to design alternatives to conventional treatment strategies, due to the increase in resistance to standard antibacterials. In this study, we present a comparative analysis of chokepoint and essentiality datasets that will provide insight into the development of new treatment regimes. We illustrate the key metabolic pathways shared between these three organisms and identify drug targets with a wide metabolic impact that are common to the three species. We demonstrate that 72% of the chokepoint enzymes are proteins essential to Mycobacterium tuberculosis. We show also that 78% of the drug targets, prioritized based on their presence in multiple paths on the metabolic network, are present in pathways shared by M. tuberculosis, M. leprae and M. abscessus, including biosynthesis of amino acids, carbohydrates, cell structures, fatty acid and lipid biosynthesis. A further 17% is found in the prioritised pathways shared between M. tuberculosis and M. abscessus. We have performed comparative structure modelling of potential drug targets identified using our analysis in order to assess druggability and demonstrate the importance of chokepoint analysis in terms of drug target identification. AUTHOR SUMMARYComputer simulation studies to design new drugs against mycobacteria

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Rational design of drug-like compounds targeting Mycobacterium marinum MelF protein

et al. 2017

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The mycobacterial mel2 locus (mycobacterial enhanced infection locus, Rv1936-1941) is Mycobacterium marinum and M. tuberculosis specific, which can withstand reactive oxygen species (ROS) and reactive nitrogen species (RNS) induced stress. A library of over a million compounds was screened using in silico virtual ligand screening (VLS) to identify inhibitors against the modeled structure of MelF protein expressed by melF of mel2 locus so that M. marinum’s ability to withstand ROS/RNS stress could be reduced. The top ranked 1000 compounds were further screened to identify 178 compounds to maximize the scaffold diversity by manually evaluating the interaction of each compound with the target site. M. marinum melF was cloned, expressed and purified as maltose binding protein (MBP)-tagged recombinant protein in Escherichia coli. After establishing the flavin dependent oxidoreductase activity of MelF (~ 84 kDa), the inhibitors were screened for the inhibition of enzyme activity of whole cell lysate (WCL) and the purified MelF. Amongst these, 16 compounds could significantly inhibit the enzyme activity of purified MelF. For the six best inhibitory compounds, the minimal inhibitory concentration (MIC) was determined to be 3.4–19.4 μM and 13.5–38.8 μM for M. marinum and M. tuberculosis, respectively. Similarly, the minimal bactericidal concentration (MBC) was determined to be 6.8–38.8 μM and 27–38.8 μM against M. marinum and M. tuberculosis, respectively. One compound each in combination with isoniazid (INH) also showed synergistic inhibitory effect against M. marinum and M. tuberculosis with no cytotoxicity in HeLa cells. Interestingly, these inhibitors did not display any non-specific protein-structure destabilizing effect. Such inhibitors targeting the anti-ROS/RNS machinery may facilitate the efficient killing of replicating and nonreplicating mycobacteria inside the host cells.

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Preliminary X-ray crystallographic analysis of ornithine acetyltransferase (Rv1653) fromMycobacterium tuberculosis

Cited by 4 publications

References 22 publications

The Molecular Structure of Ornithine Acetyltransferase from Mycobacterium tuberculosis Bound to Ornithine, a Competitive Inhibitor

The Molecular Structure of Ornithine Acetyltransferase from Mycobacterium tuberculosis Bound to Ornithine, a Competitive Inhibitor

Analysis of metabolic pathways in mycobacteria to aid drug-target identification

Rational design of drug-like compounds targeting Mycobacterium marinum MelF protein

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