Cs0720

Citrate synthase activity was determined (Sigma-Aldrich CS0720) in isolated mitochondria using spectrophotometry. Inhibitor sensitive respiratory chain complex activities were measured in 1-mL volume spectrophotometrically in the presence of 400 μM propofol or equal volume 10% intralipid or in non-exposed mitochondria and normalized to citrate synthase activity.^{23 (link),24 (link)} Rotenone-sensitive Complex I specific activity was measured in mitochondria (40 μg) with 4.8 mM⁻¹ cm⁻¹ as the extinction coefficient of NADH at 340 nm using reference wavelength of 380 nm. 2-Thenoyltrifluoroacetone-sensitive Complex II activity was determined in mitochondria (40 μg) using 19.1 mM⁻¹ cm⁻¹ as the extinction coefficient of 2,6-dichlorophenolindophenol at 600 nm. For Complexes III and IV, inhibitor-sensitive first-order rate constants were calculated in mitochondria (4 μg and 2 μg, respectively) using 18.5 mM⁻¹ cm⁻¹ as the extinction coefficient of cytochrome c at 550 nm. Oligomycin-sensitive Complex V specific activity was measured in mitochondria (40 μg) with 6.2 mM⁻¹ cm⁻¹ as the extinction coefficient of NADH at 340 nm. Rotenone-sensitive Complex I+III linked activity and antimycin A-sensitive Complex II+III linked activity were determined separately in mitochondria (40 μg) with 18.5 mM⁻¹ cm⁻¹ as the extinction coefficient of cytochrome c at 550 nm.