Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference** |
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| Authors: | Dr Jamal El Bakali Dr Michal Blaszczyk Dr Joanna C Evans Dr Jennifer A Boland Dr William J McCarthy Imam Fathoni Dr Marcio V B Dias Dr Eachan O Johnson Dr Anthony G Coyne Prof Valerie Mizrahi Prof Tom L Blundell Prof Chris Abell Dr Christina Spry |
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| Institution: | 1. Yusuf Hamied Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW UK;2. Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA UK;3. MRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DST/NRF Centre of Excellence for Biomedical TB Research & Wellcome Centre for Infectious Diseases Research in Africa, Institute of Infectious Disease and Molecular Medicine and Department of Pathology, Faculty of Health Sciences, University of Cape Town, Anzio Road, Cape Town, Observatory, 7925 South Africa;4. Research School of Biology, The Australian National University, Linnaeus Way, ACT, 2601 Australia;5. Systems Chemical Biology of Infection and Resistance Laboratory, The Francis Crick Institute, 1 Midland Road, London, NW1 1AT UK |
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| Abstract: | The coenzyme A (CoA) biosynthesis pathway has attracted attention as a potential target for much-needed novel antimicrobial drugs, including for the treatment of tuberculosis (TB), the lethal disease caused by Mycobacterium tuberculosis (Mtb). Seeking to identify inhibitors of Mtb phosphopantetheine adenylyltransferase (MtbPPAT), the enzyme that catalyses the penultimate step in CoA biosynthesis, we performed a fragment screen. In doing so, we discovered three series of fragments that occupy distinct regions of the MtbPPAT active site, presenting a unique opportunity for fragment linking. Here we show how, guided by X-ray crystal structures, we could link weakly-binding fragments to produce an active site binder with a KD <20 μM and on-target anti-Mtb activity, as demonstrated using CRISPR interference. This study represents a big step toward validating MtbPPAT as a potential drug target and designing a MtbPPAT-targeting anti-TB drug. |
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| Keywords: | Coenzyme A Drug Discovery Enzymes Fragment-Based Tuberculosis |
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