Accéder au contenu
Merck

Chemogenomics identifies acetyl-coenzyme A synthetase as a target for malaria treatment and prevention.

Cell chemical biology (2021-08-05)
Robert L Summers, Charisse Flerida A Pasaje, Joao P Pisco, Josefine Striepen, Madeline R Luth, Krittikorn Kumpornsin, Emma F Carpenter, Justin T Munro, De Lin, Andrew Plater, Avinash S Punekar, Andrew M Shepherd, Sharon M Shepherd, Manu Vanaerschot, James M Murithi, Kelly Rubiano, Aslı Akidil, Sabine Ottilie, Nimisha Mittal, A Hazel Dilmore, Madalyn Won, Rebecca E K Mandt, Kerry McGowen, Edward Owen, Chris Walpole, Manuel Llinás, Marcus C S Lee, Elizabeth A Winzeler, David A Fidock, Ian H Gilbert, Dyann F Wirth, Jacquin C Niles, Beatriz Baragaña, Amanda K Lukens
RÉSUMÉ

We identify the Plasmodium falciparum acetyl-coenzyme A synthetase (PfAcAS) as a druggable target, using genetic and chemical validation. In vitro evolution of resistance with two antiplasmodial drug-like compounds (MMV019721 and MMV084978) selects for mutations in PfAcAS. Metabolic profiling of compound-treated parasites reveals changes in acetyl-CoA levels for both compounds. Genome editing confirms that mutations in PfAcAS are sufficient to confer resistance. Knockdown studies demonstrate that PfAcAS is essential for asexual growth, and partial knockdown induces hypersensitivity to both compounds. In vitro biochemical assays using recombinantly expressed PfAcAS validates that MMV019721 and MMV084978 directly inhibit the enzyme by preventing CoA and acetate binding, respectively. Immunolocalization studies reveal that PfAcAS is primarily localized to the nucleus. Functional studies demonstrate inhibition of histone acetylation in compound-treated wild-type, but not in resistant parasites. Our findings identify and validate PfAcAS as an essential, druggable target involved in the epigenetic regulation of gene expression.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Chloroquine diphosphate salt, powder or crystals, 98.5-101.0% (EP)
Sigma-Aldrich
n-Propionyl coenzyme A lithium salt, ≥85%
Sigma-Aldrich
Anticorps monoclonal de souris anti-HA antibody produced in mouse, clone HA-7, purified from hybridoma cell culture
Sigma-Aldrich
Trichostatine A, ≥98% (HPLC), from Streptomyces sp.
Sigma-Aldrich
Sf9 Insect Cells - Novagen, Serum-free, adapted Sf9 cells
Sigma-Aldrich
Anticorps anti-acétyl-histone H4, serum, Upstate®