Figure 2. Variation of key components during the formation of erNVs. Rates are reported for catalysis only by outward facing Ec-F₁F₀.

Data are mean averages with propagated error (SEM) values from triplicate technical replicates. Unless denoted otherwise, erNVs were prepared using the standard protocol. The ratios in panels A and C are molar ratios. See Materials and Methods for experimental details. (A) Variation of the mito-CI concentration. Different amounts of mito-CI (0, 14, 28, 56, 112, 224 μg) were co-reconstituted with 29 μg Ec-F₁F₀ using the standard protocol, and rates of ATP synthesis (black) and NADH oxidation (red) determined. (B) Dependence of the rate of ATP synthesis on ubiquinone-10 concentration. Liposomes containing 0, 2.5, 10, 15, or 20 nmol Q₁₀ (mg lipid)⁻¹ were prepared and mito-CI and Ec-F₁F₀ were co-reconstituted using the standard protocol. The final Q₁₀ concentrations shown were determined using HPLC coupled to an electrochemical detector as described in Materials and Methods. (C) Dependence of ATP synthase turnover on the concentration of AOX added. AOX was added in different amounts (0, 0.04, 0.36, 0.72, 1.44, 2.89, 5.77, and 11.55 μg mL⁻¹) to 2.5 μL preformed PLs containing 143 μg mL⁻¹ outward-facing mito-CI and 128 μg mL⁻¹ outward-facing Ec-F₁F₀. (D) Dependence of the rate of ATP synthesis on lipid composition. Liposomes containing different phospholipid compositions (% (w/w); see inset table below figure) were prepared and mito-CI and Ec-F₁F₀ co-reconstituted using the standard protocol.