Fig. 3: CoA-PanAm binds to and inhibits AcAS.
From: Preclinical characterization and target validation of the antimalarial pantothenamide MMV693183
a Cellular thermal shift assay on P. falciparum lysate. Parasite lysate at 2.1 mg/ml was aliquoted and treated with 1 µM MMV693183 or metabolites thereof, or with DHA (negative control) for 30 min, followed by a 3-min incubation at different temperatures (N = 1). Protein stabilization was analyzed on a western blot (left panel) and band intensities (AcAS molecular weight = 113.8 kDa) were quantified and normalized to DMSO treatment at 37 °C (right panel). b, AcAS activity in a dose-response assay. Wild-type (T648) or mutant (M648) AcAS was immunopurified from NF54 or NF54-HGL and cT648M mutant parasite lysate, respectively, using rabbit immune serum. The activity was measured using 14C-labeled sodium acetate upon treatment with metabolites of MMV693183 and normalized to the no drug control (N = 4 for wild-type AcAS, N = 2 for mutant AcAS). c, Immunofluorescence microscopy of parasites with AcAS fused to GFP. Depicted are representative images of asexual blood-stage parasites stained with anti-GFP antibodies and DNA stained with DAPI. Scale bars, 2 µm. PanAm: MMV693183; 4’P-PanAm: 4’P-MMV693183; CoA-PanAm: CoA-MMV693183. Source data are provided as Source Data file.
