DprE2 is a molecular target of the anti-tubercular nitroimidazole compounds pretomanid and delamanid

Abstract Mycobacterium tuberculosis is one of the global leading causes of death due to a single infectious agent. Pretomanid and delamanid are new antitubercular agents that have progressed through the drug discovery pipeline. These compounds are bicyclic nitroimidazoles that act as pro-drugs, requiring activation by a mycobacterial enzyme; however, the precise mechanisms of action of the active metabolite(s) are unclear. Here, we identify a molecular target of activated pretomanid and delamanid: the DprE2 subunit of decaprenylphosphoribose-2’-epimerase, an enzyme required for the synthesis of cell wall arabinogalactan. We also provide evidence for an NAD-adduct as the active metabolite of pretomanid. Our results highlight DprE2 as a potential antimycobacterial target and provide a foundation for future exploration into the active metabolites and clinical development of pretomanid and delamanid

Structural analysis of phosphoribosyltransferase-mediated cell wall precursor synthesis in Mycobacterium tuberculosis

In Mycobacterium tuberculosis, Rv3806c is a membrane-bound phosphoribosyltransferase (PRTase) involved in cell wall precursor production. It catalyses pentosyl phosphate transfer from phosphoribosyl pyrophosphate to decaprenyl phosphate, to generate 5-phospho-β-ribosyl-1-phosphoryldecaprenol. Despite Rv3806c being an attractive drug target, structural and molecular mechanistic insight into this PRTase is lacking. Here we report cryogenic electron microscopy structures for Rv3806c in the donor- and acceptor-bound states. In a lipidic environment, Rv3806c is trimeric, creating a UbiA-like fold. Each protomer forms two helical bundles, which, alongside the bound lipids, are required for PRTase activity in vitro. Mutational and functional analyses reveal that decaprenyl phosphate and phosphoribosyl pyrophosphate bind the intramembrane and extramembrane cavities of Rv3806c, respectively, in a distinct manner to that of UbiA superfamily enzymes. Our data suggest a model for Rv3806c-catalysed phosphoribose transfer through an inverting mechanism. These findings provide a structural basis for cell wall precursor biosynthesis that could have potential for anti-tuberculosis drug development.</p

Advances in archaeomagnetic dating in Britain: New data, new approaches and a new calibration curve

yesArchaeomagnetic dating offers a valuable chronological tool for archaeological investigations, particularly for dating fired material. The method depends on the establishment of a dated record of secular variation of the Earth's magnetic field and this paper presents new and updated archaeomagnetic directional data from the UK and geomagnetic secular variation curves arising from them. The data are taken from publications from the 1950's to the present day; 422 dated entries derived from existing archaeo and geomagnetic databases are re-evaluated and 487 new directions added, resulting in 909 entries with corresponding dates, the largest collection of dated archaeomagnetic directions from a single country. An approach to improving the largest source of uncertainty, the independent dating, is proposed and applied to the British Iron Age, resulting in 145 directions from currently available databases being updated with revised ages and/or uncertainties, and a large scale reassessment of age assignments prior to inclusion into the Magnetic Moments of the Past and GEOMAGIA50 databases. From the significantly improved dataset a new archaeomagnetic dating curve for the UK is derived through the development of a temporally continuous geomagnetic field model, and is compared with previous UK archaeomagnetic dating curves and global field models. The new model, ARCH-UK.1 allows model predictions for any location in the UK with associated uncertainties. It is shown to improve precision and accuracy in archaeomagnetic dating, and to provide new insight into past geomagnetic field changes.Arts and Humanities Research Counci

Vanoxerine kills mycobacteria through membrane depolarization and efflux inhibition

Mycobacterium tuberculosis is a deadly pathogen, currently the leading cause of death worldwide from a single infectious agent through tuberculosis infections. If the End TB 2030 strategy is to be achieved, additional drugs need to be identified and made available to supplement the current treatment regimen. In addition, drug resistance is a growing issue, leading to significantly lower treatment success rates, necessitating further drug development. Vanoxerine (GBR12909), a dopamine re-uptake inhibitor, was recently identified as having anti-mycobacterial activity during a drug repurposing screening effort. However, its effects on mycobacteria were not well characterized. Herein, we report vanoxerine as a disruptor of the membrane electric potential, inhibiting mycobacterial efflux and growth. Vanoxerine had an undetectable level of resistance, highlighting the lack of a protein target. This study suggests a mechanism of action for vanoxerine, which will allow for its continued development or use as a tool compound