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. 2022 Oct 27;13(6):e02730-22. doi: 10.1128/mbio.02730-22

FIG 4.

FIG 4

MMV688766 does not directly inhibit fungal fatty acid synthase. (A) S. cerevisiae FAS complex was purified from a 3xFLAG-tagged Fas1 (JWL01) strain (right). C. albicans FAS complex purified from a 6His-3xFLAG-tagged Fas1 strain (CaLC5425) (left). Activity was assayed for activity through spectrophotometric measurement of NADPH oxidation (334 nm) in the absence (black) and presence of 20 μM (ScFAS) or 50 μM (CaFAS) MMV688766 (blue), positive-control compound 20 μM (ScFAS) or 45 μM (CaFAS) cerulenin (red), and DMSO solvent control (green). The mean of triplicate samples is presented. Error bars depict standard deviation and data are plotted using GraphPad Prism 9. (B) C. albicans FAS purified from a 6His-3xFLAG-tagged Fas1 strain (CaLC5425) grown in the presence and absence of 25 μM MMV688766 was run on a 3 to 12% blue Native PAGE gel to determine the integrity of the complex upon treatment. (C) Simplified fungal sphingolipid biosynthetic pathway demonstrating the inhibition modes of cerulenin and myriocin. The pathway was adapted and modified from reference 37.