Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference

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Date

2023

Authors

El Bakali, J.
Blaszczyk, M.
Evans, J.C.
Boland, J.A.
McCarthy, W.J.
Fathoni, I.
Dias, M.V.B.
Johnson, E.O.
Coyne, A.G.
Mizrahi, V.

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Angewandte Chemie International Edition, 2023; 62(17):e202300221-1-e202300221-8

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Jamal El Bakali, Michal Blaszczyk, Joanna C. Evans, Jennifer A. Boland, William J. McCarthy, Imam Fathoni, Marcio V. B. Dias, Eachan O. Johnson, Anthony G. Coyne, Valerie Mizrahi, Tom L. Blundell, Chris Abell, and Christina Spry

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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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Communications

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© 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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