Solubilization and reconstitution of the Na+-motive NADH oxidase activity from the marine bacterium Vibrio alginolyticus

AbstractThe Na+-motive NADH oxidase activity from Vibrio alginolyticus was extracted with octylglucoside and reconstituted into liposomes by dilution. On the addition of NADH, the reconstituted proteoliposomes generated Δψ (inside positive) and ΔpH (inside alkaline) in the presence of a proton conductor CCCP, and accumulated Na+ in the presence of valinomycin. These results indicate that the NADH oxidase activity, reconstituted in opposite orientation, leads to the generation of an eletrochemical potential of Na+ by the influx of Na+