2. Academic Validation
  3. THPP target assignment reveals EchA6 as an essential fatty acid shuttle in mycobacteria

THPP target assignment reveals EchA6 as an essential fatty acid shuttle in mycobacteria

  • Nat Microbiol. 2016 Jan 18;1:15006. doi: 10.1038/nmicrobiol.2015.6.
Jonathan A G Cox 1,  Katherine A Abrahams 1,  Carlos Alemparte 2,  Sonja Ghidelli-Disse 3,  Joaquín Rullas 2,  Iñigo Angulo-Barturen 2,  Albel Singh 1,  Sudagar S Gurcha 1,  Vijayashankar Nataraj 1,  Stephen Bethell 1,  Modesto J Remuiñán 2,  Lourdes Encinas 2,  Peter J Jervis 1,  Nicholas C Cammack 2,  Apoorva Bhatt 1,  Ulrich Kruse 3,  Marcus Bantscheff 3,  Klaus Fütterer 1,  David Barros 2,  Lluis Ballell 2,  Gerard Drewes 3,  Gurdyal S Besra 1
Affiliations

Affiliations

  • 1 Institute of Microbiology and Infection, School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
  • 2 Diseases of the Developing World, GlaxoSmithKline, Severo Ochoa 2, Tres Cantos, Madrid 28760, Spain.
  • 3 Cellzome, Meyerhofstrasse 1, Heidelberg 69117, Germany.
PMID: 27571973 DOI: 10.1038/nmicrobiol.2015.6
Abstract

Phenotypic screens for bactericidal compounds against drug-resistant tuberculosis are beginning to yield novel inhibitors. However, reliable target identification remains challenging. Here, we show that tetrahydropyrazo[1,5-a]pyrimidine-3-carboxamide (THPP) selectively pulls down EchA6 in a stereospecific manner, instead of the previously assigned target Mycobacterium tuberculosis MmpL3. While homologous to mammalian enoyl-coenzyme A (CoA) hydratases, EchA6 is non-catalytic yet essential and binds long-chain acyl-CoAs. THPP inhibitors compete with CoA-binding, suppress mycolic acid synthesis, and are bactericidal in a mouse model of chronic tuberculosis Infection. A point mutation, W133A, abrogated THPP-binding and increased both the in vitro minimum inhibitory concentration and the in vivo effective dose 99 in mice. Surprisingly, EchA6 interacts with selected Enzymes of fatty acid synthase II (FAS-II) in Bacterial two-hybrid assays, suggesting essentiality may be linked to feeding long-chain fatty acids to FAS-II. Finally, our data show that spontaneous resistance-conferring mutations can potentially obscure the actual target or alternative targets of small molecule inhibitors.

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