Figure 6.

Metal-catalyzed oxidation of mitochondrial membrane proteins and inactivation of ANT. Mitochondria were incubated with 1 mM vanadyl sulfate and 1 mM H₂O₂, followed by either membrane isolation or ANT activity measurements. (A) Immunochemical detection of ANT without oxidation (lane 1) and with oxidation (lane 2). (B) Increase of total mitochondrial membrane protein carbonyls after oxidation. Carbonyls were determined spectrophotometrically. (C) Loss of ANT activity caused by metal-catalyzed oxidation. Whole mitochondria were incubated with various concentrations of the oxidants (vanadyl/H₂O₂ = 1:1) before ANT activity measurements. For both B and C, values are the mean ± SD of four independent determinations that used 10-day-old flies hatched on different dates.