Figure 5.

Inhibition of the electron transfer activity of mitoNEET by the pioglitazone analog NL-1. MitoNEET (containing 10 μm [2Fe-2S] clusters) was preincubated with increasing concentrations of NL-1 (from 0–250 μm) at room temperature for 30 min under aerobic conditions, followed by addition of FMN (0.1 μm) and flavin reductase (0.1 μm). NADH (50 μm) was then added to the incubation solutions to initiate the reaction. The amount of the reduced mitoNEET [2Fe-2S] clusters in the incubation solutions was measured from the different absorption at absorption peaks of 455 nm and 420 nm and plotted as function of the reaction time after injection of NADH. The data are representative of three independent experiments.