The use of NAC fatty acid mimics to assay individual proteins in the FASII cycle.
(a) Incubation of (E)-2-decenoyl-NAC and PaFabV in the presence of NADH leads to formation of decanoyl-NAC. The consumption of NADH can be monitored by fluorescence.
(b) Incubation of 3-hydroxydecanoyl-NAC with PaFabA that converts it into (E)-2-decenoyl-NAC. The formation of the double bond can be monitored by direct UV measurement.
(c) The log-dose response of N42FTA in the direct (UV) assay for PaFabA, each point is an average of three measurements and the error bars represent the standard deviation.